Comparative analysis of biochemical parameters in the blood of malaria patients: correlation with treatment response to artemether-lumefantrine combination therapy

Comparative analysis of biochemical parameters in the blood of malaria patients: correlation with treatment response to artemether-lumefantrine combination therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Comparative analysis of biochemical parameters in the blood of malaria patients: correlation with treatment response to artemether-lumefantrine combination therapy Miebaka Jamabo, Michael. O Okpara, Nkemjika Amadi, Tamunodiepriye Oliseh, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8673475/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Background Malaria remains a major health concern in many parts of the world. Globally, about 260 million malaria cases and 600,000 malaria-related deaths, caused by different Plasmodium spp ., are recorded annually. Biochemical markers play a crucial role in the diagnosis or prognosis of malaria, and the blood levels of the associated biomarkers could be altered upon infection or post-treatment with antimalarials. This study aimed to investigate the impact of artemether-lumefantrine combination therapy on the serum levels of biochemical markers in malaria patients. Method Blood samples were collected from 30 malaria patients and matched with those from 20 healthy control subjects. Parasitaemia and blood levels of selected liver function, kidney function, and inflammation markers were measured in malaria patients and healthy control subjects. The malaria patients were treated with the standard dose of artemether-lumefantrine combination therapy. After recovery from malaria, parasitaemia and blood levels of the selected markers were measured again. Results During infection with P. falciparum , serum ALT and AST were reduced, parasitaemia and serum CRP were increased, while serum urea, creatinine, and neopterin remained statistically unchanged when compared with their levels in healthy control subjects. After treatment with artemether-lumefantrine combination therapy, serum creatinine and CRP levels were reduced, AST was increased, while parasitaemia, ALT, urea, and neopterin levels remained statistically unchanged when compared with their levels in malaria patients pre-treatment. Conclusion Taken together, our data demonstrated that infection with P. falciparum can modulate the serum levels of key hepatic, renal, and inflammatory markers. Furthermore, treatment with artemether-lumefantrine combination therapy is effective in reversing some of the pathological changes caused by P. falciparum in malaria patients. malaria ALT AST urea creatinine CRP neopterin artemether lumefantrine Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Background Malaria is a major health problem in many parts of the world, with about 263 million cases and 597,000 deaths recorded in 2023 alone [ 1 ]. In humans, malaria is a disease caused by either Plasmodium falciparum , vivax , malariae , ovale , or knowlesi , which are transmitted by the bite of an infected female Anopheles mosquito [ 2 ]. The most frequent clinical manifestation of malaria infection is fever with other symptoms like headache, nausea, vomiting, and/or diarrhoea, which usually appear 7–14 days following the bite from an infected mosquito [ 3 ]. Malaria is a common tropical disease prevalent among people living in countries like Nigeria, Uganda, Mali, Tanzania, and Ghana [ 4 ], [ 5 ]. Nigeria is reported to have the highest burden of malaria globally [ 6 ], [ 7 ]. This is mostly because of poor living conditions, climate change, and a lack of proper healthcare infrastructure for the prognosis, diagnosis, and treatment of malaria [ 5 ], [ 8 ], [ 9 ], [ 10 ]. Biomarkers play a crucial role in diagnosis, prognosis, therapeutics, and general understanding of disease mechanisms [ 11 ]. Diagnostic biomarkers are helpful for early detection of diseases, prognostic biomarkers provide information on possible outcomes, while therapeutic markers serve as targets for therapy [ 12 ], [ 13 ]. Malaria infection significantly alters biochemical markers of liver and kidney function. The levels of liver enzymes, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), were elevated in malaria patients compared to healthy individuals [ 14 ], [ 15 ], [ 16 ]. Similarly, kidney function markers such as creatinine, urea, and urinary protein were also significantly increased in malaria patients compared to healthy individuals [ 14 ], [ 15 ]. Inflammatory markers like C-reactive protein (CRP), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were elevated in complicated malaria cases, though their individual diagnostic accuracy is limited [ 17 ]. In a Zambian study on children, neopterin levels were shown to correlate with severe malaria but were inversely correlated to hemoglobin (Hb) levels in Ghanaian children with different severities of malaria [ 18 ], [ 19 ]. Neopterin is an immune activation marker produced by activated monocytes and macrophages and is essential for monitoring inflammatory diseases [ 20 ], [ 21 ]. The levels of these haematological parameters pre-, during, and post-treatment of malaria could serve as biomarkers for the diagnosis and prognosis of malaria. Artemisinin-based combination therapies (ACTs) have been fundamental in the global effort to reduce the burden of malaria and are currently used as first-line malaria therapy in endemic countries [ 22 ], [ 23 ]. Older and popular antimalarial drugs like chloroquine lost efficacy due to widespread resistance [ 24 ], [ 25 ], [ 26 ]. Consequently, ACTs gained prevalence due to their broad stage specificity of action, as well as their lower chances of drug resistance [ 27 ], [ 28 ]. Six ACTs are currently recommended by the World Health Organization (WHO) for treating malaria cases worldwide: i) artesunate + amodiaquine (AS + AQ), ii) artemether + lumefantrine (AL), iii) artesunate + sulfadoxine-pyrimethamine (AS + SP), iv) artesunate + mefloquine (AS + MQ), v) artesunate + pyronaridine (AS + PY) and vi) dihydroartemisinin + piperaquine (DHA + PPQ) [ 22 ], [ 29 ]. Artemether-lumefantrine and artesunate-amodiaquine are adopted in treatment guidelines for uncomplicated P. falciparum malaria in the majority of sub-Saharan African countries. Studies conducted from 2010–2020 on the efficacy of ACTs used in sub-Saharan Africa revealed overall high malaria treatment success for artemether-lumefantrine and artesunate-amodiaquine, even above the WHO threshold value, suggesting that, presently, there is no need for a change in treatment policy in Sub-Saharan African countries [ 30 ], [ 31 ], [ 32 ], [ 23 ]. With the high influx of malaria patients recorded in Rivers State University in Nigeria and the consistent use of the WHO-approved artemether-lumefantrine combination therapy, this study aims to investigate the possible effects of artemether-lumefantrine on various biochemical markers, as well as explore these markers as a tool for further exploring better prognosis, diagnosis, and treatment of malaria. Materials and Methods Ethical statement Ethical approval was received from the Rivers State University Research and Development Centre before the commencement of the study. The study was approved in compliance with the Ethical Principles for Medical Research Involving Human Subjects adopted by the World Medical Association Declaration of Helsinki [33]. Ethical clearance was approved on 22 nd April, 2025 with approval number – RSU-REC/2025/FS/0002. Permission was obtained from the hospital used and participants were adequately informed about the purpose and procedure of the study. Written consent was obtained from each participant before sample collection. Confidentiality of participants’ data was strictly maintained throughout the study. Study design and selection of participants This is a cross-sectional study comparing malaria patients (before and after treatment) with healthy controls. The clinical records of the participants were reviewed, and any participant whose record showed a history or recent diagnosis of hepatitis B and C, HIV/AIDS, liver disease, tuberculosis, kidney disease, or any chronic or infectious disease was excluded from this study. Pregnant participants were also excluded from this study. These exclusions were necessary to avoid including participants with underlying health conditions, which could influence the outcome of this study. After reviewing the clinical records of the participants, thirty (30) clinically confirmed and randomly selected male and female outpatients attending the clinic in Rivers State University Medical Centre were enrolled for this study. All laboratory investigations were carried out in Rivers State University Medical Centre Laboratory and Image Diagnostics. These centres are located in Port Harcourt City, Rivers State, Nigeria. Age-matched asymptomatic/non-malaria male and female subjects (n=20) constituted the control subjects. All participants in this study were students (male and female) from Rivers State University between the ages of 18-25. Study area The research was carried out in Rivers State University Medical Centre, Port Harcourt, Rivers State, Nigeria. The medical centre serves as the primary healthcare facility for students and staff of the institution and was selected because of its accessibility to the target population and availability of laboratory support for malaria diagnosis. The study was conducted between July and September 2025. This period coincides with the rainy season in the south-south region of Nigeria. The rainy season spans the period of March/April to October/November [34], [35]. This is a period when breeding of Anopheles mosquitoes is at its peak. Consequently, mosquito bites and transmission of Plasmodium spp. are prevalent during this period. Collection and preparation of blood specimens Blood specimens were collected by venipuncture from the study participants using 10 mL capacity disposable syringes. A total of 8 mL of blood was transferred into 15 mL Falcon tubes and centrifuged at 2000 x g for 10 min to obtain serum. The serum samples were stored at 4°C until the biochemical tests were performed. The remaining 2 mL was transferred into an EDTA anti-coagulating tube for malaria parasite test. For the malaria patients, blood samples were collected before and after malaria treatment. All samples were properly labelled with patient identifier code, data and time of collection. Malaria parasite density test From the blood samples collected, thin blood films were made, air-dried and fixed with methanol. These were stained with Giemsa stain for 10 min. The slides were viewed under the microscope using a x100 objective lens. Both parasitised and non-parasitised red blood cells were counted in several fields, and the parasitaemia level was estimated and recorded. Parasite density was calculated, and results were expressed as the number of parasites per microliter of blood. %Parasitaemia was calculated using the formula: Drug administration Malaria patients received the standard dose of artemether–lumefantrine according to the National Malaria Treatment Guidelines. Dosage was determined based on patient body weight, and adherence was ensured under clinical supervision [36]. Biochemical analyses ALT and AST liver function tests ALT and AST activities were measured using Spectrum Diagnostics Liquizyme (1 + 1) reagent kits (REF 263 001-003[ALT], REF 259 001-003 [AST], following protocols aligned with the International Federation of Clinical Chemistry (IFCC) guidelines [37], [38], [39]. Spectrophotometric measurements were performed at 340 nm using a UV-visible spectrophotometer with a 1 cm light path at 37°C. ALT catalyses the transamination of L-alanine and α-ketoglutarate to form pyruvate and glutamate. Pyruvate reacts with NADH in the presence of LDH to form lactate and NAD⁺. AST catalyses the transamination of L-aspartate and 2-oxoglutarate to form oxaloacetate and glutamate. Oxaloacetate is reduced to malate by MDH, with concurrent oxidation of NADH to NAD⁺. Working reagents were prepared by mixing equal volumes of Reagent 1 and Reagent 2 for each assay. For the ALT assay, 1000 µL of Reagent 1 (buffer containing LDH and EDTA), 250 µL of Reagent 2 (2-ketoglutarate and NADH), and 100 µL of serum sample were pipetted into test tubes. For the AST assay, 1000 µL of working reagent (Tris buffer pH 7.8, L-aspartate, LDH ≥800 U/L, MDH ≥400 U/L, EDTA, and NADH 0.18 mmol/L) and 100 µL of serum sample were pipetted into test tubes. The contents of the test tubes were mixed gently and incubated at 37°C. Absorbance readings were taken after 1 min and then every minute for 3 min. The average change in absorbance per minute (ΔA/min) was calculated, and enzyme activities were computed using the IFCC formulae. Urea Urea concentrations in serum were measured using the Modified Urease-Berthlot Colorimetric Method with Spectrum Liquizyme reagents [40]. This enzymatic assay quantifies urea based on its hydrolysis by urease and subsequent colorimetric detection of the resulting ammonia. Urea is enzymatically hydrolysed by urease to produce ammonia and carbon dioxide. The released ammonia reacts with sodium salicylate and hypochlorite in an alkaline medium to form a green-coloured complex. The intensity of the colour, measured at 578 nm, is directly proportional to the urea concentration. In this study, the reaction was set up by adding 1.0 mL of R1 buffer (phosphate buffer pH 8.0, sodium salicylate, sodium nitroprusside, EDTA) to each cuvette, followed by 50 µL of R2 enzyme (urease >6000 U/L), and 10 µL of sample or standard. The contents of the cuvette were mixed and incubated at 37°C for 3 min. For colour development, 200 µL of R3 alkaline reagent (sodium hydroxide, sodium hypochlorite) was added, and the mixture was incubated at 37°C for 5 min. Absorbance was measured at 578 nm against a reagent blank, and urea concentration in the samples was determined. Creatinine Creatinine levels were quantitatively determined in human serum using the Jaffe colorimetric kinetic method [41], [42]. This assay relies on the reaction between creatinine and alkaline picrate, forming a red-coloured complex. The intensity of the colour, measured spectrophotometrically at 492 nm, is directly proportional to the creatinine concentration. A working reagent was prepared by mixing equal volumes of picric acid reagent (R1, 17.5 mmol/L) and alkaline reagent (R2, sodium hydroxide 0.29 mol/L). Briefly, 1.0 mL of working reagent was added to cuvettes containing either 100 µL of standard or sample. Absorbance readings were taken at 30 seconds (A1) and 90 seconds (A2) post-reaction initiation. ΔA (A2 – A1) was calculated for both the sample and standard, and creatinine concentration was determined. C-reactive protein Serum concentrations of C-reactive protein (CRP) were quantified using a high-sensitivity enzyme-linked immunosorbent assay (hsCRP ELISA) purchased from BioCheck Inc. (Cat. No. BC-1119). This assay is based on a solid-phase sandwich ELISA technique utilising monoclonal and polyclonal antibodies specific to CRP. The assay was performed according to the manufacturer’s instructions. Briefly, microtiter wells were pre-coated with mouse monoclonal anti-CRP antibodies. Diluted serum samples (1:100) were incubated with a horseradish peroxidase-conjugated goat anti-CRP antibody. CRP molecules formed a sandwich complex between the immobilised and enzyme-linked antibodies. After incubation at 25°C for 45 min, the wells of the plate were washed with water to remove unbound antibody. Tetramethylbenzidine (TMB) substrate was added and incubated at 25°C for 20 min, during which a blue colour developed. The reaction was stopped with 1N HCl, changing the colour from blue to yellow. Absorbance was measured at 450 nm using a UV/visible spectrophotometer. CRP concentration in the serum sample was directly proportional to the colour intensity and was calculated from a standard curve. Neopterin This study employed a quantitative enzyme-linked immunosorbent assay (ELISA) to determine Human Neopterin (NP) concentrations in serum samples. Neopterin levels were measured using the Human Neopterin ELISA Kit (Nanjing Aoqing Medical Equipment Co., Ltd, Cat. No. JX-76673A1) according to the manufacturer’s instructions. The assay was based on a sandwich ELISA technique. Neopterin in the sample bound to antibodies pre-coated on the microplate. After washing, an enzyme-linked antibody specific to neopterin was added. Following incubation and washing, a substrate solution was added, producing a colorimetric reaction proportional to the neopterin concentration. All reagents and samples were prepared according to the kit instructions. Briefly, 50 µL of standards or samples was added to designated wells, followed by 50 µL of enzyme conjugate to each well. The plate was incubated at 37°C for 60 min before the wells were washed 5 times with the wash buffer. A volume of 50 µL of substrate solution was added to each well, and the plate was incubated at 25°C for 15 min in the dark. The intensity of the blue colouration formed was proportional to the amount of neopterin present in the serum sample. A volume of 50 µL of stop solution was added to each well to terminate the reaction, changing the colour from blue to yellow. Absorbance was measured at 450 nm within 15 min using a UV/visible spectrophotometer. Neopterin concentrations were calculated from a standard curve. Statistical analyses The graphs and statistical analyses performed in this study were plotted using GraphPad Prism software version 8 (GraphPad Software Inc., CA, USA). Box and whisker plots, showing minimum to maximum values, were used to represent the serum levels of the parameters/markers. The Comparisons for each parameter/marker between the control subjects and patients (pre- and post-treatment) were performed using ordinary one-way ANOVA with Tukey’s multiple comparison test. For comparisons between male and female genders, bar graphs showing mean±SEM, were used to represent the serum levels of the parameters/markers. The comparisons for each parameter/marker between the control subjects and patients (pre- and post-treatment) in male and female participants were performed using two-way ANOVA with Tukey’s multiple comparison test. For all statistical analyses in this study, * P <0.05, ** P <0.01, *** P <0.001, and **** P <0.0001 were considered significant. The Pearson correlation analyses in this study were performed using Microsoft Excel software version 16 (Microsoft Inc., WA, USA). Results A total of 50 participants (male and female) enrolled in this study (Table 1), comprising 30 students with confirmed P. falciparum malaria and 20 healthy, matched controls. Blood samples were obtained from all participants. Malaria patients provided paired samples, before and after malaria treatment, while each healthy control subject provided their blood sample only once. All malaria cases were confirmed microscopically. There was a significant increase in parasitaemia in malaria patients (pre-treatment) compared to the control subjects (Figure 1A). The increased parasitaemia in malaria patients was reduced slightly upon treatment with artemether-lumefantrine (Figure 1A). Similar trends were observed when parasitaemia in male and female malaria patients pre-treatment were compared with parasitaemia in either the control subjects or malaria patients post-treatment with artemether-lumefantrine (Figure 1B). Table 1. Demographic and parasitaemia characteristics of study participants Characteristic Controls (n = 20) Malaria Patients (n = 30) Gender 5 Male, 15 Female 12 Male, 18 Female Age range 18 – 25 years 18 – 25 years Population Undergraduate students Undergraduate students Drug administered – Artemether-lumefantrine Parasitaemia (%) 0 – 2% (very low) Before treatment: 0 – 23% After treatment: 0 – 21% In the present study, the serum levels of ALT and AST were significantly reduced in malaria patients (pre-treatment) compared to ALT and AST serum levels in healthy control subjects (Figures 2A and B). Upon treatment with artemether-lumefantrine combination therapy, there was an increase in serum ALT level compared to the level before treatment. Although, this increase was not statistically significant. Serum AST level was significantly increased after treatment with artemether-lumefantrine combination therapy compared to serum AST level before treatment (Figures 2A and B). Gender-based analysis showed that the male malaria patients, but not the female patients, had a significant reduction in Serum ALT levels when compared to the healthy control subjects (Figure 3A). Across both genders, serum AST levels were significantly reduced in malaria patients pre-treatment compared to both the healthy control subjects and malaria patients post-treatment (Figure 3B). The serum levels of the kidney function markers, urea and creatinine, were statistically unchanged in malaria patients before treatment compared to healthy subject controls (Figure 4A and B). However, upon treatment of the malaria patients with artemether-lumefantrine combination therapy, serum creatinine level, but not urea, was significantly reduced (Figure 4A and B). Statistically, there is no difference in serum urea levels between healthy control subjects and malaria patients pre- or post-treatment, regardless of gender (Figure 5A). However, the serum level of creatinine was significantly reduced in both genders after treatment with artemether-lumefantrine combination therapy (Figure 5B). Serum CRP levels were markedly increased in malaria patients pre-treatment compared to healthy control subjects. Interestingly, serum CRP levels in the malaria patients significantly reduced after treatment with artemether-lumefantrine combination therapy (Figure 6A. Contrarily, neopterin concentrations showed no statistically significant alterations across all groups (Figure 6B). CRP, but not neopterin, showed a clear trend of infection-induced upregulation followed by post-treatment decline (Figure 6). Similar patterns were observed across both genders (Figure 7A and B). Correlation analyses (Tables 2 and 3) were performed to explore the interrelationships among all the markers measured in this study, before and after treatment. During the pre-treatment phase, most biomarkers under investigation in this study showed weak or negligible correlations. However, after treatment with artemether-lumefantrine combination therapy, a few notable associations emerged (Table 3). The most pronounced was a strong positive correlation between ALT and AST (r=0.74). A moderate positive correlation was observed between AST and creatinine (r = 0.54), A similar, albeit weaker, relationship between ALT and creatinine (r=0.35) was also noted. Other parameters, including CRP, neopterin, and urea, exhibited weak or no significant correlations with one another either before or after treatment. The correlations between parasitaemia and most biochemical markers was also weak, except a mild positive correlation with urea before treatment (r=0.36) Table 2. Correlation analysis between biochemical parameters pre-treatment with artemether-lumefantrine combination therapy. Pre-treatment ALT AST Urea Creatinine CRP Neopterin Parasitaemia ALT 1 AST 0.071 1 Urea -0.176 0.079 1 Creatinine 0.012 -0.198 0.095 1 CRP 0.117 0.087 -0.027 -0.205 1 Neopterin -0.056 0.358 0.183 0.038 0.040 1 Parasitaemia 0.085 0.000 0.363 0.086 -0.184 0.061 1 Table 3. Correlation analysis between biochemical parameters post-treatment with artemether-lumefantrine combination therapy. Post-treatment ALT AST Urea Creatinine CRP Neopterin Parasitaemia ALT 1 AST 0.743 1 Urea 0.175 0.278 1 Creatinine 0.351 0.538 0.259 1 CRP -0.189 -0.054 0.102 0.210 1 Neopterin 0.154 0.137 0.235 0.047 0.001 1 Parasitaemia -0.162 -0.016 0.155 0.128 -0.074 -0.153 1 The correlation coefficients are numerical values ranging from -1 to +1. Values near +1 indicate a strong positive relationship. Values near -1 indicate a strong negative relationship. Values near 0 indicate little to no linear relationship. Discussion Malaria continues to be a major public health issue, with 263 million cases and 597,000 deaths recorded in 2023 [1]. Current malaria treatment strategies primarily focus on parasite clearance and symptom relief without considering individual variations in biochemical and immune response. Also, the emergence of drug-resistant plasmodium strains and variability in patient responses to antimalarial drugs highlight the need for a deeper understanding of the disease's pathophysiology [43], [44]. Biochemical, oxidative, and immune/inflammatory markers are known to be altered during malaria infection, but their role in predicting drug response remains unclear [45], [46]. Understanding the interplay of these markers in malaria patients and their correlation with treatment response can provide valuable insights to optimise therapeutic outcomes, identify biomarkers for monitoring disease progression and possibly reduce drug resistance. Serum levels of liver and kidney function biomarkers are reported to be altered during and after malaria infection [14], [15], [47]. The changes in ALT and AST observed in this study are consistent with hepatic involvement during malaria infection. The malaria parasite is known to affect many organs, and liver dysfunction is an associated feature in malaria [48], [49]. The alterations in uncomplicated malaria are typically transient, resolving as parasites are cleared and hepatic regeneration begins [50], [51]. The non-significant post-treatment increase in ALT in our cohort suggests partial restoration of hepatic function within the seven-day observation period. This aligns with findings that normalisation may take up to two weeks post-infection clearance [51], [52]. Also, in a retrospective study done between 2010 and 2017 on returning travellers who tested positive for malaria, no abnormalities were seen for all liver enzymes [53]. The reduction of serum creatinine suggests an improved renal handling following parasite clearance (Figure 4). Severe malaria is usually associated with acute kidney injury (AKI) [54] and acute renal failure can occur in 1% of severe cases. This is more common in older populations [55], however, it could occur in children [56], [57]. The findings from this study corroborate earlier reports that uncomplicated malaria seldom results in acute kidney injury [58]. The reduction in serum creatinine level post-treatment further supports improved renal clearance following parasite elimination. Further research with a larger and more diverse population set must be conducted to clarify sex-dependent variations in malaria pathophysiology. Malaria is known to trigger a strong inflammatory response, leading to the release of cytokines, which are responsible for symptoms like fever and headache [59]. C-reactive protein (CRP) is an established acute-phase protein produced in response to proinflammatory cytokines such as IL-6 and is usually released during malaria infection [60], [61]. Our findings support previous reports identifying CRP as a sensitive biomarker for malaria diagnosis and severity assessment [62], [61], [17]. Neopterin, a macrophage-derived molecule reflecting Th1-type immune activation, was markedly elevated in malaria patients, consistent with heightened cellular immunity during active infection [21]. According to some studies, neopterin levels are usually highest in first-time malaria patients as well as in severe P. falciparum cases [63], [64]. The significant reduction in CRP after therapy confirms its utility as a prognostic biomarker in monitoring treatment response and resolution of inflammation and malaria [62], [61], [17]. Previous studies have shown a correlation between parasite burden, gender, and some biochemical markers [65], [66], [67]. The absence of strong associations at this stage likely reflects the complex and multifactorial nature of the host response to Plasmodium infection, where hepatic, renal, and immune systems are variably affected depending on parasite load and individual immune competence [68] After treatment with artemether-lumefantrine combination therapy, notable associations emerged. The strong positive correlation between ALT and AST (r=0.74) indicates concurrent normalisation of hepatocellular enzymes and consistent hepatic recovery after therapy. These finding aligns with the well-established biochemical interdependence of these transaminases, which both serve as sensitive indicators of hepatocellular integrity [69]. The moderate positive correlation between AST and creatinine (r = 0.54) may reflect systemic improvement in metabolic and renal function as inflammation subsides post-treatment. A similar, albeit weaker, relationship between ALT and creatinine (r=0.35) suggests partial hepatic–renal linkage in the recovery process [47], [70]. Other parameters, including CRP, neopterin, and urea, exhibited weak or no significant correlations with one another either before or after treatment. This pattern implies that inflammatory and immune markers fluctuate independently of hepatic and renal parameters, possibly due to differences in their regulation and turnover rates. CRP and neopterin are both acute-phase reactants influenced by cytokine activity and macrophage stimulation, which may not directly be in parallel with enzymatic or excretory biomarkers. Interestingly, the weak negative correlations between CRP and some hepatic or renal indices post-treatment could indicate an inverse relationship between inflammation resolution and organ stress. However, this can only be ascertained with a larger sample size. The lack of strong correlations between parasitaemia and most biochemical markers, except a mild positive correlation with urea before treatment (r=0.36), suggests that parasite density alone does not fully account for biochemical perturbations. Instead, host factors such as immune response intensity and oxidative stress likely contribute significantly to biochemical changes during infection. Overall, the correlation patterns reveal that, while malaria exerts widespread metabolic influence, organ-specific responses are only loosely interlinked. The emergence of stronger correlations post-treatment underscores the coordinated normalisation of physiological systems following parasite clearance and therapeutic recovery. Conclusion This study demonstrated that uncomplicated P. falciparum malaria in the student population of Rivers State University was characterized by slightly significant but reversible dysfunction in both hepatic and renal systems, accompanied by a robust inflammatory response. Artemether-lumefantrine therapy was effective in reducing parasite burden and facilitating the restoration of organ function. Growing evidence suggests that biological sex is a variable that can impact immune response, drug metabolism, physiology, and consequently, the progression of a disease [71]. P. falciparum malaria infection showed a sex-specific host-pathogen interaction in the liver, similar to other hepatotropic parasitic and viral pathogens [72], [73]. This means males were more likely to experience a delayed detection of blood-stage parasites in a controlled human malaria infection trial with P. falciparum compared to females. However, the parasite replication rates upon blood stage emergence showed no significant differences between the sexes [74]. Other studies have shown that due to economic and socio-behavioural factors, there was an increased risk of males getting the infection [75], [76] but females were able to clear asymptomatic infections faster than men [77]. The absence of significant gender-based differences in our study may be attributable to our homogeneous study population of students/young adults. This suggests that in uncomplicated malaria, the pathophysiological processes, treatment response, and recovery process are not markedly influenced by gender. Although the study was limited by sample size and follow-up duration, it provides valuable baseline data from a university setting in Nigeria. Future studies should incorporate longitudinal sampling, oxidative stress indices, and cytokine profiles to deepen understanding of malaria-associated biochemical pathways. In conclusion, this study provides a comprehensive profile of organ dysfunction and systemic inflammation in uncomplicated P. falciparum malaria and demonstrates the effectiveness of artemether-lumefantrine in reversing these pathological changes. The comparable response across genders supports the universal application of this treatment regimen in similar demographic groups. Abbreviations AST aspartate aminotransferase ALT alanine aminotransferase CRP C-reactive protein AL artemether-lumefantrine ELISA enzyme-linked immunosorbent assay Declarations Ethical approval was received from the Rivers State University Research and Development Centre before the commencement of the study. The study was approved in compliance with the Ethical Principles for Medical Research Involving Human Subjects adopted by the World Medical Association Declaration of Helsinki [ 33 ]. Ethical clearance was approved on 22nd April, 2025 with approval number – RSU-REC/2025/FS/0002. Permission was obtained from the hospital used and participants were adequately informed about the purpose and procedure of the study. Written consent was obtained from each participant before sample collection. Confidentiality of participants’ data was strictly maintained throughout the study. Funding No funds, grants, or other support was received for conducting this study Author Contribution M.J and M.O.O: conceptualization, study design, and methodology.M.J: Drafted the initial manuscript.M.O.O: performed statistical analysis, critically reviewed and edited the manuscript.O.J: Oversaw drug dissemination, supervised participant follow-up, and sample collection.N.A and T.O: Enrolled participants and collected clinical data.B.B: Conducted sample collection and performed biochemical assays.All authors have reviewed and approved the final version of the manuscript Data Availability All data supporting the findings of this study are available within the paper or on request References WHO World malaria report 2024. Accessed: Nov. 10, 2025. [Online]. Available: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2024 Kho S et al (2022) Diagnostic performance of a 5-plex malaria immunoassay in regions co-endemic for Plasmodium falciparum, P. vivax, P. knowlesi, P. malariae and P. ovale, Sci. Rep. , vol. 12, no. 1, p. 7286, May 10.1038/s41598-022-11042-w Islam MR, Dhar PS, Rahman MM (2023) Recently outbreak of malaria in current world: species, etiology, life cycle, transmission, symptoms, vaccination, diagnostic tests, treatment, and complications, Int. J. Surg. , vol. 109, no. 2, pp. 175–177, Feb. 10.1097/JS9.0000000000000165 Howick VM et al (Aug. 2019) The Malaria Cell Atlas: Single parasite transcriptomes across the complete Plasmodium life cycle. Science 365(6455):eaaw2619. 10.1126/science.aaw2619 Chimezie RO (2020) Malaria Hyperendemicity: The Burden and Obstacles to Eradication in Nigeria. J Biosci Med 08(11):165–178. 10.4236/jbm.2020.811015 Shekarau E et al (Jun. 2024) Severe malaria intervention status in Nigeria: workshop meeting report. Malar J 23(1):177. 10.1186/s12936-024-05001-1 WHO World malaria report 2023. Accessed: Nov. 11, 2025. [Online]. Available: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2023 WHO Report on malaria in Nigeria 2022 | WHO | Regional Office for Africa. Accessed: Mar. 12, 2025. [Online]. Available: https://www.afro.who.int/countries/nigeria/publication/report-malaria-nigeria-2022 Ayanlade A et al (Dec. 2022) Climate change engenders a better Early Warning System development across Sub-Saharan Africa: The malaria case. Resour Environ Sustain 10:100080. 10.1016/j.resenv.2022.100080 Aderoyeje TG, Erhuanga OG (2024) A Review on the Exploration of Genomic Approaches to Malaria Prevention and Treatment in Nigeria, J. Appl. Sci. Environ. Manag. , vol. 28, no. 9, pp. 2937–2941, Sep. 10.4314/jasem.v28i9.40 Manning L, Davis TME (2013) The Mechanistic, Diagnostic and Prognostic Utility of Biomarkers in Severe Malaria, Biomark. Med. , vol. 7, no. 3, pp. 363–380, Jun. 10.2217/bmm.13.50 Foko LPK et al (2022) The spectrum of clinical biomarkers in severe malaria and new avenues for exploration, Virulence , vol. 13, no. 1, pp. 634–653, Dec. 10.1080/21505594.2022.2056966 Hunter DJ, Losina E, Guermazi A, Burstein D, Lassere MN, Kraus V (May 2010) A Pathway and Approach to Biomarker Validation and Qualification for Osteoarthritis Clinical Trials. Curr Drug Targets 11(5):536–545 Ogbadoyi E, Tsado R (2009) Renal and Hepatic Dysfunction in Malaria Patients in Minna, North Central Nigeria, Nov. Accessed: Mar. 15, 2025. [Online]. Available: https://www.semanticscholar.org/paper/Renal-and-Hepatic-Dysfunction-in-Malaria-Patients-Ogbadoyi-Tsado/eb81837108481004f9f296447592af32aaa31726 Oyewole I, Senusie S, Mansaray M (2010) Plasmodium Falciparum-Induced Kidney and Liver Dysfunction in Malaria Patients in Freetown, Sierra Leone, Sierra Leone J. Biomed. Res. , vol. 2, no. 1, pp. 70–74, Jul. 10.4314/sljbr.v2i1.56611 Singh G, Urhekar A, Singh R, Maheshwari UM, Samant P (2015) Alteration in biochemical parameters in malaria patients. Plasmodium falciparum vs. Plasmodium vivax, Accessed: Mar. 15, 2025. [Online]. Available: https://www.semanticscholar.org/paper/Alteration-in-biochemical-parameters-in-malaria-vs.-Singh-Urhekar/ad6358a5192eb5417f58a2ff2cafdf3eaa7fee3a Hashmi F, Aqeel S, Zuberi UF, Khan W (Sep. 2023) A systematic review and meta-analysis of inflammatory biomarkers associated with malaria infection and disease severity. Cytokine 169:156305. 10.1016/j.cyto.2023.156305 Biemba G, Gordeuk VR, Thuma PE, Mabeza GF, Weiss G (Jan. 1998) Prolonged macrophage activation and persistent anaemia in children with complicated malaria. Trop Med Int Health 3(1):60–65. 10.1046/j.1365-3156.1998.00168.x Awandare GA et al (2006) Increased Levels of Inflammatory Mediators in Children with Severe Plasmodium falciparum Malaria with Respiratory Distress, J. Infect. Dis. , vol. 194, no. 10, pp. 1438–1446, Nov. 10.1086/508547 Gil-Gómez R, Blasco-Alonso J, Sánchez-Yáñez P, Rosa-Camacho V, Manso GM (2017) Niveles de neopterina y síndrome de respuesta inflamatoria sistémica en pacientes críticos pediátricos, An. Pediatría , vol. 87, no. 6, pp. 343–349, Dec. 10.1016/j.anpedi.2016.11.011 Blankson SO et al (Jan. 2023) High Plasma Levels of Neopterin Are Associated with Increased Mortality among Children with Severe Malaria in Benin. Diagnostics 13(3):528. 10.3390/diagnostics13030528 Arya A, Kojom Foko LP, Chaudhry S, Sharma A, Singh V (2021) Artemisinin-based combination therapy (ACT) and drug resistance molecular markers: A systematic review of clinical studies from two malaria endemic regions – India and sub-Saharan Africa, Int. J. Parasitol. Drugs Drug Resist. , vol. 15, pp. 43–56, Apr. 10.1016/j.ijpddr.2020.11.006 Marwa K et al (2022) Therapeutic efficacy of artemether-lumefantrine, artesunate-amodiaquine and dihydroartemisinin-piperaquine in the treatment of uncomplicated Plasmodium falciparum malaria in Sub-Saharan Africa: A systematic review and meta-analysis, PLOS ONE , vol. 17, no. 3, p. e0264339, Mar. 10.1371/journal.pone.0264339 Ursos LMB, Roepe PD (2002) Chloroquine resistance in the malarial parasite, Plasmodium falciparum , Med. Res. Rev. , vol. 22, no. 5, pp. 465–491, Sep. 10.1002/med.10016 Baird JK (Nov. 2004) Chloroquine Resistance in Plasmodium vivax . Antimicrob Agents Chemother 48(11):4075–4083. 10.1128/AAC.48.11.4075-4083.2004 Chinappi M, Via A, Marcatili P, Tramontano A (Nov. 2010) On the Mechanism of Chloroquine Resistance in Plasmodium falciparum. PLoS ONE 5(11):e14064. 10.1371/journal.pone.0014064 Ashley EA et al (2014) Spread of Artemisinin Resistance in Plasmodium falciparum Malaria, N. Engl. J. Med. , vol. 371, no. 5, pp. 411–423, Jul. 10.1056/NEJMoa1314981 Blasco B, Leroy D, Fidock DA (Aug. 2017) Antimalarial drug resistance: linking Plasmodium falciparum parasite biology to the clinic. Nat Med 23(8):917–928. 10.1038/nm.4381 WHO WHO: World malaria report, 2013 - Google Scholar. Accessed: Oct. 15, 2025. [Online]. Available: https://scholar.google.com/scholar_lookup?title=World%20Malaria%20Report%202019&publication_year=2019&author=WHO Byakika-Kibwika P, Lamorde M, Mayanja-Kizza H, Khoo S, Merry C, Van Geertruyden J-P (2011) Artemether-Lumefantrine Combination Therapy for Treatment of Uncomplicated Malaria: The Potential for Complex Interactions with Antiretroviral Drugs in HIV-Infected Individuals, Malar. Res. Treat. , vol. pp. 1–5, Apr. 2011. 10.4061/2011/703730 Zwang J et al (Dec. 2012) Comparing changes in haematologic parameters occurring in patients included in randomized controlled trials of artesunate-amodiaquine vs single and combination treatments of uncomplicated falciparum in sub-Saharan Africa. Malar J 11(1):25. 10.1186/1475-2875-11-25 Schramm B et al (Dec. 2013) Efficacy of artesunate-amodiaquine and artemether-lumefantrine fixed-dose combinations for the treatment of uncomplicated Plasmodium falciparum malaria among children aged six to 59 months in Nimba County, Liberia: an open-label randomized non-inferiority trial. Malar J 12(1):251. 10.1186/1475-2875-12-251 WMA, (2000) World Medical Association Declaration of Helsinki: Ethical Principles for Medical Research Involving Human Subjects Revised October 7, 2000, HIV Clin. Trials , vol. 2, no. 1, pp. 92–95, Feb. 2001. 10.1310/GTFR-2DRX-M6YE-ELXR Rawlings A, Ehiorobo J, Izinyon C (2020) Assessment of Rainfall Seasonality and Distribution Pattern in the South-South Region of Nigeria, vol. 17, pp. 4040–4059, Apr Ibebuchi CC, Abu I-O (May 2023) Rainfall variability patterns in Nigeria during the rainy season. Sci Rep 13(1):7888. 10.1038/s41598-023-34970-7 Olugbenga MA, Obot O-OA, Oluwatoyin HA (2018) Effect of Artemisinin Based Combination Therapy on the Liver and Kidney of Patient Attending University Health Centre. Niger J Pharm Appl Sci Res 7(2):28–32 Schumann G, Klauke R (Jan. 2003) New IFCC reference procedures for the determination of catalytic activity concentrations of five enzymes in serum: preliminary upper reference limits obtained in hospitalized subjects. Clin Chim Acta Int J Clin Chem 327:1–2. 10.1016/s0009-8981(02)00341-8 Ceriotti F et al (2010) Common reference intervals for aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) in serum: results from an IFCC multicenter study, Clin. Chem. Lab. Med. , vol. 48, no. 11, pp. 1593–1601, Nov. 10.1515/CCLM.2010.315 Agarwal AK, Katiyar GD, Khan S, Chaudhary BC, Singh A, Sharma M (May 2020) Evaluation of liver function test in malaria positive cases in tertiary care hospital of Bareilly. Int J Community Med Public Health 7(6):2193–2202. 10.18203/2394-6040.ijcmph20202470 Mišić IDR, Mitić SS, Kostić DA, Tošić SB, Pecev-Marinković ET, Miletić AS (2021) Kinetic-spectrophotometric approach to the modified Berthelot procedure for serum urea determination, Chem. Pap. , vol. 75, no. 2, pp. 565–574, Feb. 10.1007/s11696-020-01315-x Marakala V, Arnadi Ramachandrayya ASSS, Mala M, Kumar A (Oct. 2012) SERUM CREATININE ASSAY: ENZYMATIC VS KINETIC JAFFE’S METHOD. J Evol Med Dent Sci 1:258–264. 10.14260/jemds/54 Vani K, Bodepudi SR, Praveen V Study of Creatinine Estimation by Enzymatic and Kinetic Jaffe’s Method White NJ (2004) Antimalarial drug resistance, J. Clin. Invest. , vol. 113, no. 8, pp. 1084–1092, Apr. 10.1172/JCI21682 Abebe W, Woldesenbet D (Aug. 2025) A review on molecular markers of Plasmodium falciparum. J Parasit Dis. 10.1007/s12639-025-01840-0 Vasquez M, Zuniga M, Rodriguez A (Nov. 2021) Oxidative Stress and Pathogenesis in Malaria. Front Cell Infect Microbiol 11:768182. 10.3389/fcimb.2021.768182 Chandrashekhar VN et al (2022) Impact of oxidative stress in response to malarial infection during pregnancy: Complications, histological changes, and pregnancy outcomes, Trop. Parasitol. , vol. 12, no. 1, p. 21, Jun. 10.4103/tp.TP_18_20 Singh G, URHEKAR A, Singh R, MAHESHWARI U, SAMANT P, Al A (2015) Alteration in biochemical parameters in malaria patients. Plasmodium falciparum vs. Plasmodium vivax, J. Microbiol. Antimicrob. AGENTS , vol. 1, Jan Kochar DK et al (Jul. 2003) Hepatocyte dysfunction and hepatic encephalopathy in Plasmodium falciparum malaria. QJM 96(7):505–512. 10.1093/qjmed/hcg091 Olusola AO (2018) Effect of Plasmodium Falciparum on Liver Function Parameters of Children in Akoko Area of Ondo State, Nigeria. J Diagn 5(1):1–5. 10.18488/journal.98.2018.51.1.5 Reuling IJ et al (Oct. 2018) Liver Injury in Uncomplicated Malaria is an Overlooked Phenomenon: An Observational Study. eBioMedicine 36:131–139. 10.1016/j.ebiom.2018.09.018 Tangpukdee N et al (2006) Minor liver profile dysfunctions in Plasmodium vivax, P. malariae and P. ovale patients and normalization after treatment. Korean J Parasitol 44(4):295. 10.3347/kjp.2006.44.4.295 Odedra A, Woodford J, Chalon S, Barber BE, McCarthy JS (2022) Posttreatment Transaminase Elevations in Controlled Human Malaria Infection and Naturally Acquired Malaria, J. Infect. Dis. , vol. 225, no. 5, pp. 926–927, Mar. 10.1093/infdis/jiab502 Cheaveau J et al (Jun. 2019) The Impact of Malaria on Liver Enzymes: A Retrospective Cohort Study (2010–2017). Open Forum Infect Dis 6(6):ofz234. 10.1093/ofid/ofz234 Trang TTM et al (Nov. 1992) Acute Renal Failure in Patients with Severe Falciparum Malaria. Clin Infect Dis 15(5):874–880. 10.1093/clind/15.5.874 Mishra SK, Das BS (Jul. 2008) Malaria and Acute Kidney Injury. Semin Nephrol 28(4):395–408. 10.1016/j.semnephrol.2008.04.007 Conroy AL et al (Apr. 2016) Acute Kidney Injury Is Common in Pediatric Severe Malaria and Is Associated With Increased Mortality. Open Forum Infect Dis 3(2):ofw046. 10.1093/ofid/ofw046 Afolayan FM et al (Nov. 2020) Acute Kidney Injuries in Children with Severe Malaria: A comparative study of diagnostic criteria based on serum cystatin C and creatinine levels. Sultan Qaboos Univ Med J 20(4):e312–e317. 10.18295/squmj.2020.20.04.006 Katsoulis O, Georgiadou A, Cunnington AJ (Apr. 2021) Immunopathology of Acute Kidney Injury in Severe Malaria. Front Immunol 12:651739. 10.3389/fimmu.2021.651739 Farrington L et al (Dec. 2017) Both inflammatory and regulatory cytokine responses to malaria are blunted with increasing age in highly exposed children. Malar J 16(1):499. 10.1186/s12936-017-2148-6 Mackintosh CL, Beeson JG, Marsh K (Dec. 2004) Clinical features and pathogenesis of severe malaria. Trends Parasitol 20(12):597–603. 10.1016/j.pt.2004.09.006 Wilairatana P et al (Nov. 2021) C-reactive protein as an early biomarker for malaria infection and monitoring of malaria severity: a meta-analysis. Sci Rep 11(1):22033. 10.1038/s41598-021-01556-0 Bhardwaj N, Ahmed M, Sharma S, Nayak A, Anvikar A, Pande V (2019) C-reactive protein as a prognostic marker of Plasmodiumfalciparum malaria severity. J Vector Borne Dis 56(2):122. 10.4103/0972-9062.263727 Brown AE, Webster HK, Teja-Isavadharm P, Keeratithakul D (Oct. 1990) Macrophage activation in falciparum malaria as measured by neopterin and interferon-gamma. Clin Exp Immunol 82(1):97–101. 10.1111/j.1365-2249.1990.tb05410.x Te Witt R et al (Dec. 2010) Neopterin and procalcitonin are suitable biomarkers for exclusion of severe Plasmodium falciparum disease at the initial clinical assessment of travellers with imported malaria. Malar J 9(1):255. 10.1186/1475-2875-9-255 Nagaraj N, Berwal PK, Srinivas A, Prakash P, Ramesh MS, Berwal A (2018) Correlation of hepatorenal dysfunction in pediatric malaria. Trop Parasitol 8(2):83–87. 10.4103/tp.TP_2_16 Plewes K et al (Mar. 2014) Correlation of biomarkers for parasite burden and immune activation with acute kidney injury in severe falciparum malaria. Malar J 13:91. 10.1186/1475-2875-13-91 Ojo RJ, Jonathan IG, Adams MD, Gyebi G, Longdet IY (Oct. 2022) Renal and hepatic dysfunction parameters correlate positively with gender among patients with recurrent malaria cases in Birnin Kebbi, Northwest Nigeria. Egypt J Intern Med 34(1):77. 10.1186/s43162-022-00164-2 Ali MO, Tamomh AG, Abdulrahman NA, Ibrahim NS, Eltayeb MA, Osman FI (Sep. 2025) Exploring the Effect of Malaria on Biochemical Parameters Among Sudanese Patients with Complicated Falciparum Malaria. Clin Lab 71(9). 10.7754/Clin.Lab.2025.250208 Khan JM, Hindoliya BS, Paul A, Shahrukh M (2023) A Comparative Study Of Liver Enzymes In The Individuals With Malarial Infection And Normal Healthy Individuals, Eurasian J. Anal. Chem. , vol. 18, no. 1, pp. 12–16, Dec Godse RR (2013) Hematological and biochemical evaluation in malaria patients with clinical correlation, Accessed: Nov. 22, 2025. [Online]. Available: https://www.semanticscholar.org/paper/Hematological-and-biochemical-evaluation-in-malaria-Rr/13e5817eec55076385af5261e5ea023f01434d00 Lockard RD, Wilson ME, Rodríguez NE (2019) Sex-Related Differences in Immune Response and Symptomatic Manifestations to Infection with Leishmania Species, J. Immunol. Res. , vol. pp. 1–14, Jan. 2019. 10.1155/2019/4103819 Ruggieri A, Gagliardi MC, Anticoli S (Oct. 2018) Sex-Dependent Outcome of Hepatitis B and C Viruses Infections: Synergy of Sex Hormones and Immune Responses? Front Immunol 9:2302. 10.3389/fimmu.2018.02302 Brown R, Goulder P, Matthews PC (2022) Sexual Dimorphism in Chronic Hepatitis B Virus (HBV) Infection: Evidence to Inform Elimination Efforts. Wellcome Open Res 7:32. 10.12688/wellcomeopenres.17601.3 Duncombe CJ et al Sex-Based Differences to Plasmodium Infection in the Placebo Arms of Controlled Human Malaria Infection (CHMI) Trials in Malaria-Naïve Populations: A Pooled Analysis, Jan. 13, 2025, Social Science Research Network, Rochester, NY : 5087394. 10.2139/ssrn.5087394 Moon JJ, Cho S-Y (2001) Incidence patterns of vivax malaria in civilians residing in a high-risk county of Kyonggi-do (Province), Republic of Korea, Korean J. Parasitol. , vol. 39, no. 4, pp. 293–299, Dec. 10.3347/kjp.2001.39.4.293 Pathak S et al (2012) Age-Dependent Sex Bias in Clinical Malarial Disease in Hypoendemic Regions, PLOS ONE , vol. 7, no. 4, p. e35592, Apr. 10.1371/journal.pone.0035592 Briggs J et al (2020) Sex-based differences in clearance of chronic Plasmodium falciparum infection, eLife , vol. 9, p. e59872, Oct. 10.7554/eLife.59872 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 15 Feb, 2026 Reviews received at journal 13 Feb, 2026 Reviewers agreed at journal 13 Feb, 2026 Reviews received at journal 06 Feb, 2026 Reviewers agreed at journal 06 Feb, 2026 Reviewers agreed at journal 05 Feb, 2026 Reviewers invited by journal 05 Feb, 2026 Editor assigned by journal 04 Feb, 2026 Submission checks completed at journal 29 Jan, 2026 First submitted to journal 22 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8673475","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":587642712,"identity":"0e909683-056a-4025-8a5c-1545bd9bf08b","order_by":0,"name":"Miebaka Jamabo","email":"data:image/png;base64,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","orcid":"","institution":"Rivers State University","correspondingAuthor":true,"prefix":"","firstName":"Miebaka","middleName":"","lastName":"Jamabo","suffix":""},{"id":587642713,"identity":"d62af114-9eb6-41d7-95fc-23fa5ab5e87e","order_by":1,"name":"Michael. O Okpara","email":"","orcid":"","institution":"Rhodes University","correspondingAuthor":false,"prefix":"","firstName":"Michael.","middleName":"O","lastName":"Okpara","suffix":""},{"id":587642714,"identity":"877a291b-7b32-46e0-8ec1-8611937ec8a2","order_by":2,"name":"Nkemjika Amadi","email":"","orcid":"","institution":"Rivers State University","correspondingAuthor":false,"prefix":"","firstName":"Nkemjika","middleName":"","lastName":"Amadi","suffix":""},{"id":587642715,"identity":"fa2ad5ae-ba37-4035-bc6d-0f2761c06d97","order_by":3,"name":"Tamunodiepriye Oliseh","email":"","orcid":"","institution":"Rivers State University","correspondingAuthor":false,"prefix":"","firstName":"Tamunodiepriye","middleName":"","lastName":"Oliseh","suffix":""},{"id":587642716,"identity":"7267c04e-d859-47fd-a424-b3fc60d40e96","order_by":4,"name":"Oraibi Jamabo","email":"","orcid":"","institution":"Rivers State University","correspondingAuthor":false,"prefix":"","firstName":"Oraibi","middleName":"","lastName":"Jamabo","suffix":""},{"id":587642717,"identity":"d6efb756-4ca6-4e25-aa59-2305a9d1dc74","order_by":5,"name":"Baridomamene B. Barivole","email":"","orcid":"","institution":"Rivers State University","correspondingAuthor":false,"prefix":"","firstName":"Baridomamene","middleName":"B.","lastName":"Barivole","suffix":""}],"badges":[],"createdAt":"2026-01-22 22:38:30","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8673475/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8673475/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102376049,"identity":"1cce705c-77b0-4e7b-95db-ec9ee63be9c9","added_by":"auto","created_at":"2026-02-11 05:26:03","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1131567,"visible":true,"origin":"","legend":"\u003cp\u003e(A) Parasitaemia in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy. (B) Parasitaemia in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy for male and female participants.\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/d3bf83792995ce0d35221b14.png"},{"id":102376299,"identity":"6c764ed5-efd3-43a5-8e86-937ee81e5477","added_by":"auto","created_at":"2026-02-11 05:26:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1038376,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) ALT and (B) AST levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/c257aa708c992fa66a7ec7a3.png"},{"id":102376296,"identity":"0d69b75d-019a-4a18-93ad-88dfd2627f3d","added_by":"auto","created_at":"2026-02-11 05:26:55","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":904738,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) ALT and (B) AST levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy for male and female participants.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/ed9883e459408eba2f8c6785.png"},{"id":102376025,"identity":"2b8824d3-68ee-4e3d-a326-6ae142fe8110","added_by":"auto","created_at":"2026-02-11 05:25:59","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1160718,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) urea and (B) creatinine levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/5ba171aae96a003db877d25c.png"},{"id":102376000,"identity":"91f343de-17bc-4264-aa9e-4ea352fa7aa6","added_by":"auto","created_at":"2026-02-11 05:25:49","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":983797,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) urea and (B) creatinine levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy for male and female participants.\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/c583b08fe0b330819db47aef.png"},{"id":102376063,"identity":"6d6458c8-52ec-4465-9a0d-00e92e353c72","added_by":"auto","created_at":"2026-02-11 05:26:07","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1120257,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) CRP and (B) neopterin levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/e2daece82163f68b37e60ea8.png"},{"id":102376180,"identity":"b7e36a23-d18d-4245-be64-865cf1711ee1","added_by":"auto","created_at":"2026-02-11 05:26:31","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":958731,"visible":true,"origin":"","legend":"\u003cp\u003eSerum (A) CRP and (B) neopterin levels in healthy control subjects and malaria patients pre- and post-treatment with artemether-lumefantrine combination therapy for male and female participants.\u003c/p\u003e","description":"","filename":"image7.png","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/e176ab18934112d33b33c789.png"},{"id":102376332,"identity":"444d1a19-8b98-424d-b291-9a1fcc80cea5","added_by":"auto","created_at":"2026-02-11 05:27:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8198700,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8673475/v1/bb807bda-6255-441e-94af-bceeb9b11c3e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparative analysis of biochemical parameters in the blood of malaria patients: correlation with treatment response to artemether-lumefantrine combination therapy","fulltext":[{"header":"Background","content":"\u003cp\u003eMalaria is a major health problem in many parts of the world, with about 263\u0026nbsp;million cases and 597,000 deaths recorded in 2023 alone [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. In humans, malaria is a disease caused by either \u003cem\u003ePlasmodium falciparum\u003c/em\u003e, \u003cem\u003evivax\u003c/em\u003e, \u003cem\u003emalariae\u003c/em\u003e, \u003cem\u003eovale\u003c/em\u003e, or \u003cem\u003eknowlesi\u003c/em\u003e, which are transmitted by the bite of an infected female Anopheles mosquito [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The most frequent clinical manifestation of malaria infection is fever with other symptoms like headache, nausea, vomiting, and/or diarrhoea, which usually appear 7\u0026ndash;14 days following the bite from an infected mosquito [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Malaria is a common tropical disease prevalent among people living in countries like Nigeria, Uganda, Mali, Tanzania, and Ghana [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Nigeria is reported to have the highest burden of malaria globally [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. This is mostly because of poor living conditions, climate change, and a lack of proper healthcare infrastructure for the prognosis, diagnosis, and treatment of malaria [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBiomarkers play a crucial role in diagnosis, prognosis, therapeutics, and general understanding of disease mechanisms [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Diagnostic biomarkers are helpful for early detection of diseases, prognostic biomarkers provide information on possible outcomes, while therapeutic markers serve as targets for therapy [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Malaria infection significantly alters biochemical markers of liver and kidney function. The levels of liver enzymes, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), were elevated in malaria patients compared to healthy individuals [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Similarly, kidney function markers such as creatinine, urea, and urinary protein were also significantly increased in malaria patients compared to healthy individuals [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Inflammatory markers like C-reactive protein (CRP), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were elevated in complicated malaria cases, though their individual diagnostic accuracy is limited [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In a Zambian study on children, neopterin levels were shown to correlate with severe malaria but were inversely correlated to hemoglobin (Hb) levels in Ghanaian children with different severities of malaria [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Neopterin is an immune activation marker produced by activated monocytes and macrophages and is essential for monitoring inflammatory diseases [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The levels of these haematological parameters pre-, during, and post-treatment of malaria could serve as biomarkers for the diagnosis and prognosis of malaria.\u003c/p\u003e \u003cp\u003eArtemisinin-based combination therapies (ACTs) have been fundamental in the global effort to reduce the burden of malaria and are currently used as first-line malaria therapy in endemic countries [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Older and popular antimalarial drugs like chloroquine lost efficacy due to widespread resistance [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Consequently, ACTs gained prevalence due to their broad stage specificity of action, as well as their lower chances of drug resistance [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Six ACTs are currently recommended by the World Health Organization (WHO) for treating malaria cases worldwide: i) artesunate\u0026thinsp;+\u0026thinsp;amodiaquine (AS\u0026thinsp;+\u0026thinsp;AQ), ii) artemether\u0026thinsp;+\u0026thinsp;lumefantrine (AL), iii) artesunate\u0026thinsp;+\u0026thinsp;sulfadoxine-pyrimethamine (AS\u0026thinsp;+\u0026thinsp;SP), iv) artesunate\u0026thinsp;+\u0026thinsp;mefloquine (AS\u0026thinsp;+\u0026thinsp;MQ), v) artesunate\u0026thinsp;+\u0026thinsp;pyronaridine (AS\u0026thinsp;+\u0026thinsp;PY) and vi) dihydroartemisinin\u0026thinsp;+\u0026thinsp;piperaquine (DHA\u0026thinsp;+\u0026thinsp;PPQ) [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Artemether-lumefantrine and artesunate-amodiaquine are adopted in treatment guidelines for uncomplicated \u003cem\u003eP. falciparum\u003c/em\u003e malaria in the majority of sub-Saharan African countries. Studies conducted from 2010\u0026ndash;2020 on the efficacy of ACTs used in sub-Saharan Africa revealed overall high malaria treatment success for artemether-lumefantrine and artesunate-amodiaquine, even above the WHO threshold value, suggesting that, presently, there is no need for a change in treatment policy in Sub-Saharan African countries [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e With the high influx of malaria patients recorded in Rivers State University in Nigeria and the consistent use of the WHO-approved artemether-lumefantrine combination therapy, this study aims to investigate the possible effects of artemether-lumefantrine on various biochemical markers, as well as explore these markers as a tool for further exploring better prognosis, diagnosis, and treatment of malaria.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003ch2\u003eEthical statement\u003c/h2\u003e\n\u003cp\u003eEthical approval was received from the Rivers State University Research and Development Centre before the commencement of the study. The study was approved in compliance with the Ethical Principles for Medical Research Involving Human Subjects adopted by the World Medical Association Declaration of Helsinki [33]. Ethical clearance was approved on 22\u003csup\u003end\u003c/sup\u003e April, 2025 with approval number \u0026ndash; RSU-REC/2025/FS/0002. Permission was obtained from the hospital used and participants were adequately informed about the purpose and procedure of the study. Written consent was obtained from each participant before sample collection. Confidentiality of participants\u0026rsquo; data was strictly maintained throughout the study.\u003c/p\u003e\n\u003ch2\u003eStudy design and selection of participants\u003c/h2\u003e\n\u003cp\u003eThis is a cross-sectional study comparing malaria patients (before and after treatment) with healthy controls.\u0026nbsp;The clinical records of the participants were reviewed, and any participant whose record showed a history or recent diagnosis of hepatitis B and C, HIV/AIDS, liver disease, tuberculosis, kidney disease, or any chronic or infectious disease was excluded from this study. Pregnant participants were also excluded from this study. These exclusions were necessary to avoid including participants with underlying health conditions, which could influence the outcome of this study. After reviewing the clinical records of the participants, thirty (30) clinically confirmed and randomly selected male and female outpatients attending the clinic in Rivers State University Medical Centre were enrolled for this study. All laboratory investigations were carried out in Rivers State University Medical Centre Laboratory and Image Diagnostics. These centres are located in Port Harcourt City, Rivers State, Nigeria. Age-matched asymptomatic/non-malaria male and female subjects (n=20) constituted the control subjects. All participants in this study were students (male and female) from Rivers State University between the ages of 18-25.\u003c/p\u003e\n\u003ch2\u003eStudy area\u003c/h2\u003e\n\u003cp\u003eThe research was carried out in Rivers State University Medical Centre, Port Harcourt, Rivers State, Nigeria. The medical centre serves as the primary healthcare facility for students and staff of the institution and was selected because of its accessibility to the target population and availability of laboratory support for malaria diagnosis. The study was conducted between July and September 2025. This period coincides with the rainy season in the south-south region of Nigeria. The rainy season spans the period of March/April to October/November [34], [35]. This is a period when breeding of Anopheles mosquitoes is at its peak. Consequently, mosquito bites and transmission of \u003cem\u003ePlasmodium spp.\u003c/em\u003e are prevalent during this period.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eCollection and preparation of blood specimens\u003c/h2\u003e\n\u003cp\u003eBlood specimens were collected by venipuncture from the study participants using 10 mL capacity disposable syringes. A total of 8 mL of blood was transferred into 15 mL Falcon tubes and centrifuged at 2000 x g for 10 min to obtain serum. The serum samples were stored at 4\u0026deg;C until the biochemical tests were performed. The remaining 2 mL was transferred into an EDTA anti-coagulating tube for malaria parasite test. For the malaria patients, blood samples were collected before and after malaria treatment. All samples were properly labelled with patient identifier code, data and time of collection.\u003c/p\u003e\n\u003ch2\u003eMalaria parasite density test\u003c/h2\u003e\n\u003cp\u003eFrom the blood samples collected, thin blood films were made, air-dried and fixed with methanol. These were stained with Giemsa stain for 10 min. The slides were viewed under the microscope using a x100 objective lens. Both parasitised and non-parasitised red blood cells were counted in several fields, and the parasitaemia level was estimated and recorded. Parasite density was calculated, and results were expressed as the number of parasites per microliter of blood.\u003c/p\u003e\n\u003cp\u003e%Parasitaemia was calculated using the formula:\u003c/p\u003e\n\u003cp\u003e\u003cimg width=\"316\" height=\"33\" src=\"data:image/png;base64,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\" alt=\"image\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eDrug administration\u003c/h2\u003e\n\u003cp\u003eMalaria patients received the standard dose of artemether\u0026ndash;lumefantrine according to the National Malaria Treatment Guidelines. Dosage was determined based on patient body weight, and adherence was ensured under clinical supervision [36].\u003c/p\u003e\n\u003ch2\u003eBiochemical analyses\u003c/h2\u003e\n\u003ch3\u003e\u003cem\u003eALT and AST liver function tests\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eALT and AST activities were measured using Spectrum Diagnostics Liquizyme (1 + 1) reagent kits (REF 263 001-003[ALT], REF 259 001-003 [AST], following protocols aligned with the International Federation of Clinical Chemistry (IFCC) guidelines [37], [38], [39]. Spectrophotometric measurements were performed at 340 nm using a UV-visible spectrophotometer with a 1 cm light path at 37\u0026deg;C. ALT catalyses the transamination of L-alanine and \u0026alpha;-ketoglutarate to form pyruvate and glutamate. Pyruvate reacts with NADH in the presence of LDH to form lactate and NAD⁺. AST catalyses the transamination of L-aspartate and 2-oxoglutarate to form oxaloacetate and glutamate. Oxaloacetate is reduced to malate by MDH, with concurrent oxidation of NADH to NAD⁺. Working reagents were prepared by mixing equal volumes of Reagent 1 and Reagent 2 for each assay. \u0026nbsp;For the ALT assay, 1000 \u0026micro;L of Reagent 1 (buffer containing LDH and EDTA), 250 \u0026micro;L of Reagent 2 (2-ketoglutarate and NADH), and 100 \u0026micro;L of serum sample were pipetted into test tubes. For the AST assay, 1000 \u0026micro;L of working reagent (Tris buffer pH 7.8, L-aspartate, LDH \u0026ge;800 U/L, MDH \u0026ge;400 U/L, EDTA, and NADH 0.18 mmol/L) and 100 \u0026micro;L of serum sample were pipetted into test tubes. The contents of the test tubes were mixed gently and incubated at 37\u0026deg;C. Absorbance readings were taken after 1 min and then every minute for 3 min. The average change in absorbance per minute (\u0026Delta;A/min) was calculated, and enzyme activities were computed using the IFCC formulae.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e\u003cem\u003eUrea\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eUrea concentrations in serum were measured using the Modified Urease-Berthlot Colorimetric Method with Spectrum Liquizyme reagents [40]. This enzymatic assay quantifies urea based on its hydrolysis by urease and subsequent colorimetric detection of the resulting ammonia. Urea is enzymatically hydrolysed by urease to produce ammonia and carbon dioxide. The released ammonia reacts with sodium salicylate and hypochlorite in an alkaline medium to form a green-coloured complex. The intensity of the colour, measured at 578 nm, is directly proportional to the urea concentration. In this study, the reaction was set up by adding 1.0 mL of R1 buffer (phosphate buffer pH 8.0, sodium salicylate, sodium nitroprusside, EDTA) to each cuvette, followed by 50 \u0026micro;L of R2 enzyme (urease \u0026gt;6000 U/L), and 10 \u0026micro;L of sample or standard. The contents of the cuvette were mixed and incubated at 37\u0026deg;C for 3 min. For colour development, 200 \u0026micro;L of R3 alkaline reagent (sodium hydroxide, sodium hypochlorite) was added, and the mixture was incubated at 37\u0026deg;C for 5 min. Absorbance was measured at 578 nm against a reagent blank, and urea concentration in the samples was determined.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e\u003cem\u003eCreatinine\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eCreatinine levels were quantitatively determined in human serum using the Jaffe colorimetric kinetic method [41], [42]. This assay relies on the reaction between creatinine and alkaline picrate, forming a red-coloured complex. The intensity of the colour, measured spectrophotometrically at 492 nm, is directly proportional to the creatinine concentration. A working reagent was prepared by mixing equal volumes of picric acid reagent (R1, 17.5 mmol/L) and alkaline reagent (R2, sodium hydroxide 0.29 mol/L). \u0026nbsp;Briefly, 1.0 mL of working reagent was added to cuvettes containing either 100 \u0026micro;L of standard or sample. Absorbance readings were taken at 30 seconds (A1) and 90 seconds (A2) post-reaction initiation. \u0026Delta;A (A2 \u0026ndash; A1) was calculated for both the sample and standard, and creatinine concentration was determined.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e\u003cem\u003eC-reactive protein\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eSerum concentrations of C-reactive protein (CRP) were quantified using a high-sensitivity enzyme-linked immunosorbent assay (hsCRP ELISA) purchased from BioCheck Inc. (Cat. No. BC-1119). This assay is based on a solid-phase sandwich ELISA technique utilising monoclonal and polyclonal antibodies specific to CRP. The assay was performed according to the manufacturer\u0026rsquo;s instructions. Briefly, microtiter wells were pre-coated with mouse monoclonal anti-CRP antibodies. Diluted serum samples (1:100) were incubated with a horseradish peroxidase-conjugated goat anti-CRP antibody. CRP molecules formed a sandwich complex between the immobilised and enzyme-linked antibodies. After incubation at 25\u0026deg;C for 45 min, the wells of the plate were washed with water to remove unbound antibody. Tetramethylbenzidine (TMB) substrate was added and incubated at 25\u0026deg;C for 20 min, during which a blue colour developed. The reaction was stopped with 1N HCl, changing the colour from blue to yellow. Absorbance was measured at 450 nm using a UV/visible spectrophotometer. CRP concentration in the serum sample was directly proportional to the colour intensity and was calculated from a standard curve.\u003c/p\u003e\n\u003ch3\u003e\u003cem\u003eNeopterin\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eThis study employed a quantitative enzyme-linked immunosorbent assay (ELISA) to determine Human Neopterin (NP) concentrations in serum samples. Neopterin levels were measured using the Human Neopterin ELISA Kit (Nanjing Aoqing Medical Equipment Co., Ltd, Cat. No. JX-76673A1) according to the manufacturer\u0026rsquo;s instructions. The assay was based on a sandwich ELISA technique. Neopterin in the sample bound to antibodies pre-coated on the microplate. After washing, an enzyme-linked antibody specific to neopterin was added. Following incubation and washing, a substrate solution was added, producing a colorimetric reaction proportional to the neopterin concentration. All reagents and samples were prepared according to the kit instructions. Briefly, 50 \u0026micro;L of standards or samples was added to designated wells, followed by 50 \u0026micro;L of enzyme conjugate to each well. The plate was incubated at 37\u0026deg;C for 60 min before the wells were washed 5 times with the wash buffer. A volume of 50 \u0026micro;L of substrate solution was added to each well, and the plate was incubated at 25\u0026deg;C for 15 min in the dark. The intensity of the blue colouration formed was proportional to the amount of neopterin present in the serum sample. A volume of 50 \u0026micro;L of stop solution was added to each well to terminate the reaction, changing the colour from blue to yellow. Absorbance was measured at 450 nm within 15 min using a UV/visible spectrophotometer. Neopterin concentrations were calculated from a standard curve.\u003c/p\u003e\n\u003ch2\u003eStatistical analyses\u003c/h2\u003e\n\u003cp\u003eThe graphs and statistical analyses performed in this study were plotted using GraphPad Prism software version 8 (GraphPad Software Inc., CA, USA). Box and whisker plots, showing minimum to maximum values, were used to represent the serum levels of the parameters/markers. The Comparisons for each parameter/marker between the control subjects and patients (pre- and post-treatment) were performed using ordinary one-way ANOVA with Tukey\u0026rsquo;s multiple comparison test. For comparisons between male and female genders, bar graphs showing mean\u0026plusmn;SEM, were used to represent the serum levels of the parameters/markers. The comparisons for each parameter/marker between the control subjects and patients (pre- and post-treatment) in male and female participants were performed using two-way ANOVA with Tukey\u0026rsquo;s multiple comparison test. For all statistical analyses in this study, *\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05, **\u003cem\u003eP\u003c/em\u003e\u0026lt;0.01, ***\u003cem\u003eP\u003c/em\u003e\u0026lt;0.001, and ****\u003cem\u003eP\u003c/em\u003e\u0026lt;0.0001 were considered significant. The Pearson correlation analyses in this study were performed using Microsoft Excel software version 16 (Microsoft Inc., WA, USA).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eA total of 50 participants (male and female) enrolled in this study (Table 1), comprising 30 students with confirmed \u003cem\u003eP. falciparum\u003c/em\u003e malaria and 20 healthy, matched controls. Blood samples were obtained from all participants. Malaria patients provided paired samples, before and after malaria treatment, while each healthy control subject provided their blood sample only once. All malaria cases were confirmed microscopically. There was a significant increase in parasitaemia in malaria patients (pre-treatment) compared to the control subjects (Figure 1A). The increased parasitaemia in malaria patients was reduced slightly upon treatment with artemether-lumefantrine (Figure 1A). Similar trends were observed when parasitaemia in male and female malaria patients pre-treatment were compared with parasitaemia in either the control subjects or malaria patients post-treatment with artemether-lumefantrine (Figure 1B).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Demographic and parasitaemia characteristics of study participants\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCharacteristic\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eControls (n = 20)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMalaria Patients (n = 30)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGender\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5 Male, 15 Female\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e12 Male, 18 Female\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge range\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18 \u0026ndash; 25 years\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e18 \u0026ndash; 25 years\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eUndergraduate students\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eUndergraduate students\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrug administered\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eArtemether-lumefantrine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eParasitaemia (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0 \u0026ndash; 2% (very low)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBefore treatment: 0 \u0026ndash; 23%\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eAfter treatment: 0 \u0026ndash; 21%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eIn the present study, the serum levels of ALT and AST were significantly reduced in malaria patients (pre-treatment) compared to ALT and AST serum levels in healthy control subjects (Figures 2A and B). Upon treatment with artemether-lumefantrine combination therapy, there was an increase in serum ALT level compared to the level before treatment. Although, this increase was not statistically significant. Serum AST level was significantly increased after treatment with artemether-lumefantrine combination therapy compared to serum AST level before treatment (Figures 2A and B). \u0026nbsp;Gender-based analysis showed that the male malaria patients, but not the female patients, had a significant reduction in Serum ALT levels when compared to the healthy control subjects (Figure 3A). Across both genders, serum AST levels were significantly reduced in malaria patients pre-treatment compared to both the healthy control subjects and malaria patients post-treatment (Figure 3B).\u003c/p\u003e\n\u003cp\u003eThe serum levels of the kidney function markers, urea and creatinine, were statistically unchanged in malaria patients before treatment compared to healthy subject controls (Figure 4A and B). However, upon treatment of the malaria patients with artemether-lumefantrine combination therapy, serum creatinine level, but not urea, was significantly reduced (Figure 4A and B). Statistically, there is no difference in serum urea levels between healthy control subjects and malaria patients pre- or post-treatment, regardless of gender (Figure 5A). However, the serum level of creatinine was significantly reduced in both genders after treatment with artemether-lumefantrine combination therapy (Figure 5B).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSerum CRP levels were markedly increased in malaria patients pre-treatment compared to healthy control subjects. Interestingly, serum CRP levels in the malaria patients significantly reduced after treatment with artemether-lumefantrine combination therapy (Figure 6A. Contrarily, neopterin concentrations showed no statistically significant alterations across all groups (Figure 6B). CRP, but not neopterin, showed a clear trend of infection-induced upregulation followed by post-treatment decline (Figure 6). Similar patterns were observed across both genders (Figure 7A and B).\u003c/p\u003e\n\u003cp\u003eCorrelation analyses (Tables 2 and 3) were performed to explore the interrelationships among all the markers measured in this study, before and after treatment. During the pre-treatment phase, most biomarkers under investigation in this study showed weak or negligible correlations.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHowever, after treatment with artemether-lumefantrine combination therapy, a few notable associations emerged (Table 3). The most pronounced was a strong positive correlation between ALT and AST (r=0.74). A moderate positive correlation was observed between AST and creatinine (r = 0.54), A similar, albeit weaker, relationship between ALT and creatinine (r=0.35) was also noted. Other parameters, including CRP, neopterin, and urea, exhibited weak or no significant correlations with one another either before or after treatment. The correlations between parasitaemia and most biochemical markers was also weak, except a mild positive correlation with urea before treatment (r=0.36)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Correlation analysis between biochemical parameters pre-treatment with artemether-lumefantrine combination therapy.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"642\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre-treatment\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eALT\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eAST\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eUrea\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eCreatinine\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eCRP\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eNeopterin\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eParasitaemia\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eALT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAST\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.071\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eUrea\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.176\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0.079\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCreatinine\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e-0.198\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0.095\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCRP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.117\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0.087\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e-0.027\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e-0.205\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNeopterin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.056\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0.358\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0.183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.038\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0.040\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eParasitaemia\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.085\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0.363\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.086\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e-0.184\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.061\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Correlation analysis between biochemical parameters post-treatment with artemether-lumefantrine combination therapy.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"623\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost-treatment\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eALT\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eAST\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eUrea\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eCreatinine\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eCRP\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eNeopterin\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eParasitaemia\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eALT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAST\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.743\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eUrea\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.175\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.278\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCreatinine\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.351\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.538\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e0.259\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCRP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e-0.189\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e-0.054\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e0.102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.210\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNeopterin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e0.235\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.047\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eParasitaemia\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e-0.162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e-0.016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e0.155\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.128\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e-0.074\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e-0.153\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 112px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe correlation coefficients are numerical values ranging from -1 to +1. Values near +1 indicate a strong positive relationship. Values near -1 indicate a strong negative relationship. Values near 0 indicate little to no linear relationship.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eMalaria continues to be a major public health issue, with 263 million cases \u0026nbsp;and 597,000 deaths recorded in 2023 [1]. Current malaria treatment strategies primarily focus on parasite clearance and symptom relief without considering individual variations in biochemical and immune response. Also, the emergence of drug-resistant \u003cem\u003eplasmodium\u0026nbsp;\u003c/em\u003estrains and variability in patient responses to antimalarial drugs highlight the need for a deeper understanding of the disease's pathophysiology [43], [44]. Biochemical, oxidative, and immune/inflammatory markers are known to be altered during malaria infection, but their role in predicting drug response remains unclear [45], [46]. Understanding the interplay of these markers in malaria patients and their correlation with treatment response can provide valuable insights to optimise therapeutic outcomes, identify biomarkers for monitoring disease progression and possibly reduce drug resistance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSerum levels of liver and kidney function biomarkers are reported to be altered during and after malaria infection [14], [15], [47]. The changes in ALT and AST observed in this study are consistent with hepatic involvement during malaria infection. The malaria parasite is known to affect many organs, and liver dysfunction is an associated feature in malaria [48], [49]. The alterations in uncomplicated malaria are typically transient, resolving as parasites are cleared and hepatic regeneration begins [50], [51]. The non-significant post-treatment increase in ALT in our cohort suggests partial restoration of hepatic function within the seven-day observation period. This aligns with findings that normalisation may take up to two weeks post-infection clearance [51], [52]. Also, in a retrospective study done between 2010 and 2017 on returning travellers who tested positive for malaria, no abnormalities were seen for all liver enzymes [53].\u003c/p\u003e\n\u003cp\u003eThe reduction of serum creatinine suggests an improved renal handling following parasite clearance (Figure 4). Severe malaria is usually associated with acute kidney injury (AKI) [54] and acute renal failure can occur in 1% of severe cases. This is more common in older populations [55], however, it could occur in children [56], [57]. The findings from this study corroborate earlier reports that uncomplicated malaria seldom results in acute kidney injury [58]. The reduction in serum creatinine level post-treatment further supports improved renal clearance following parasite elimination. Further research with a larger and more diverse population set must be conducted to clarify sex-dependent variations in malaria pathophysiology.\u003c/p\u003e\n\u003cp\u003eMalaria is known to trigger a strong inflammatory response, leading to the release of cytokines, which are responsible for symptoms like fever and headache [59]. C-reactive protein (CRP) is an established acute-phase protein produced in response to proinflammatory cytokines such as IL-6 and is usually released during malaria infection [60], [61]. Our findings support previous reports identifying CRP as a sensitive biomarker for malaria diagnosis and severity assessment [62], [61], [17]. Neopterin, a macrophage-derived molecule reflecting Th1-type immune activation, was markedly elevated in malaria patients, consistent with heightened cellular immunity during active infection [21]. According to some studies, neopterin levels are usually highest in first-time malaria patients as well as in severe \u003cem\u003eP. falciparum\u003c/em\u003e cases [63], [64]. The significant reduction in CRP after therapy confirms its utility as a prognostic biomarker in monitoring treatment response and resolution of inflammation and malaria [62], [61], [17].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePrevious studies have shown a correlation between parasite burden, gender, and some biochemical markers [65], [66], [67]. The absence of strong associations at this stage likely reflects the complex and multifactorial nature of the host response to \u003cem\u003ePlasmodium\u003c/em\u003e infection, where hepatic, renal, and immune systems are variably affected depending on parasite load and individual immune competence [68]\u003c/p\u003e\n\u003cp\u003eAfter treatment with artemether-lumefantrine combination therapy, notable associations emerged. The strong positive correlation between ALT and AST (r=0.74) indicates concurrent normalisation of hepatocellular enzymes and consistent hepatic recovery after therapy. These finding aligns with the well-established biochemical interdependence of these transaminases, which both serve as sensitive indicators of hepatocellular integrity [69]. The moderate positive correlation between AST and creatinine (r = 0.54) may reflect systemic improvement in metabolic and renal function as inflammation subsides post-treatment. A similar, albeit weaker, relationship between ALT and creatinine (r=0.35) suggests partial hepatic–renal linkage in the recovery process [47], [70].\u003c/p\u003e\n\u003cp\u003eOther parameters, including CRP, neopterin, and urea, exhibited weak or no significant correlations with one another either before or after treatment. This pattern implies that inflammatory and immune markers fluctuate independently of hepatic and renal parameters, possibly due to differences in their regulation and turnover rates. CRP and neopterin are both acute-phase reactants influenced by cytokine activity and macrophage stimulation, which may not directly be in parallel with enzymatic or excretory biomarkers. Interestingly, the weak negative correlations between CRP and some hepatic or renal indices post-treatment could indicate an inverse relationship between inflammation resolution and organ stress. However, this can only be ascertained with a larger sample size.\u003c/p\u003e\n\u003cp\u003eThe lack of strong correlations between parasitaemia and most biochemical markers, except a mild positive correlation with urea before treatment (r=0.36), suggests that parasite density alone does not fully account for biochemical perturbations. Instead, host factors such as immune response intensity and oxidative stress likely contribute significantly to biochemical changes during infection.\u003c/p\u003e\n\u003cp\u003eOverall, the correlation patterns reveal that, while malaria exerts widespread metabolic influence, organ-specific responses are only loosely interlinked. The emergence of stronger correlations post-treatment underscores the coordinated normalisation of physiological systems following parasite clearance and therapeutic recovery.\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study demonstrated that uncomplicated \u003cem\u003eP. falciparum\u003c/em\u003e malaria in the student population of Rivers State University was characterized by slightly significant but reversible dysfunction in both hepatic and renal systems, accompanied by a robust inflammatory response. Artemether-lumefantrine therapy was effective in reducing parasite burden and facilitating the restoration of organ function.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGrowing evidence suggests that biological sex is a variable that can impact immune response, drug metabolism, physiology, and consequently, the progression of a disease [71]. \u003cem\u003eP.\u003c/em\u003e \u003cem\u003efalciparum\u003c/em\u003e malaria infection showed a sex-specific host-pathogen interaction in the liver, similar to other hepatotropic parasitic and viral pathogens [72], [73]. This means males were more likely to experience a delayed detection of blood-stage parasites in a controlled human malaria infection trial with \u003cem\u003eP. falciparum\u003c/em\u003e compared to females. However, the parasite replication rates upon blood stage emergence showed no significant differences between the sexes [74]. Other studies have shown that due to economic and socio-behavioural factors, there was an increased risk of males getting the infection [75], [76] but females were able to clear asymptomatic infections faster than men [77]. The absence of significant gender-based differences in our study may be attributable to our homogeneous study population of students/young adults. This suggests that in uncomplicated malaria, the pathophysiological processes, treatment response, and recovery process are not markedly influenced by gender.\u003c/p\u003e\n\u003cp\u003eAlthough the study was limited by sample size and follow-up duration, it provides valuable baseline data from a university setting in Nigeria. Future studies should incorporate longitudinal sampling, oxidative stress indices, and cytokine profiles to deepen understanding of malaria-associated biochemical pathways.\u003c/p\u003e\n\u003cp\u003eIn conclusion, this study provides a comprehensive profile of organ dysfunction and systemic inflammation in uncomplicated \u003cem\u003eP. falciparum\u003c/em\u003e malaria and demonstrates the effectiveness of artemether-lumefantrine in reversing these pathological changes. The comparable response across genders supports the universal application of this treatment regimen in similar demographic groups.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eAST\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003easpartate aminotransferase\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eALT\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ealanine aminotransferase\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCRP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eC-reactive protein\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eAL\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eartemether-lumefantrine\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eELISA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eenzyme-linked immunosorbent assay\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eEthical approval\u003c/h2\u003e \u003cp\u003ewas received from the Rivers State University Research and Development Centre before the commencement of the study. The study was approved in compliance with the Ethical Principles for Medical Research Involving Human Subjects adopted by the World Medical Association Declaration of Helsinki [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Ethical clearance was approved on 22nd April, 2025 with approval number \u0026ndash; RSU-REC/2025/FS/0002. Permission was obtained from the hospital used and participants were adequately informed about the purpose and procedure of the study. Written consent was obtained from each participant before sample collection. Confidentiality of participants\u0026rsquo; data was strictly maintained throughout the study.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eNo funds, grants, or other support was received for conducting this study\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eM.J and M.O.O: conceptualization, study design, and methodology.M.J: Drafted the initial manuscript.M.O.O: performed statistical analysis, critically reviewed and edited the manuscript.O.J: Oversaw drug dissemination, supervised participant follow-up, and sample collection.N.A and T.O: Enrolled participants and collected clinical data.B.B: Conducted sample collection and performed biochemical assays.All authors have reviewed and approved the final version of the manuscript\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll data supporting the findings of this study are available within the paper or on request\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eWHO World malaria report 2024. Accessed: Nov. 10, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2024\u003c/span\u003e\u003cspan address=\"https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2024\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKho S et al (2022) Diagnostic performance of a 5-plex malaria immunoassay in regions co-endemic for Plasmodium falciparum, P. vivax, P. knowlesi, P. malariae and P. ovale, \u003cem\u003eSci. Rep.\u003c/em\u003e, vol. 12, no. 1, p. 7286, May \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-022-11042-w\u003c/span\u003e\u003cspan address=\"10.1038/s41598-022-11042-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIslam MR, Dhar PS, Rahman MM (2023) Recently outbreak of malaria in current world: species, etiology, life cycle, transmission, symptoms, vaccination, diagnostic tests, treatment, and complications, \u003cem\u003eInt. J. Surg.\u003c/em\u003e, vol. 109, no. 2, pp. 175\u0026ndash;177, Feb. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1097/JS9.0000000000000165\u003c/span\u003e\u003cspan address=\"10.1097/JS9.0000000000000165\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHowick VM et al (Aug. 2019) The Malaria Cell Atlas: Single parasite transcriptomes across the complete \u003cem\u003ePlasmodium\u003c/em\u003e life cycle. Science 365(6455):eaaw2619. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1126/science.aaw2619\u003c/span\u003e\u003cspan address=\"10.1126/science.aaw2619\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChimezie RO (2020) Malaria Hyperendemicity: The Burden and Obstacles to Eradication in Nigeria. J Biosci Med 08(11):165\u0026ndash;178. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4236/jbm.2020.811015\u003c/span\u003e\u003cspan address=\"10.4236/jbm.2020.811015\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShekarau E et al (Jun. 2024) Severe malaria intervention status in Nigeria: workshop meeting report. Malar J 23(1):177. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s12936-024-05001-1\u003c/span\u003e\u003cspan address=\"10.1186/s12936-024-05001-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO World malaria report 2023. Accessed: Nov. 11, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2023\u003c/span\u003e\u003cspan address=\"https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2023\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO Report on malaria in Nigeria 2022 | WHO | Regional Office for Africa. Accessed: Mar. 12, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.afro.who.int/countries/nigeria/publication/report-malaria-nigeria-2022\u003c/span\u003e\u003cspan address=\"https://www.afro.who.int/countries/nigeria/publication/report-malaria-nigeria-2022\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAyanlade A et al (Dec. 2022) Climate change engenders a better Early Warning System development across Sub-Saharan Africa: The malaria case. Resour Environ Sustain 10:100080. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.resenv.2022.100080\u003c/span\u003e\u003cspan address=\"10.1016/j.resenv.2022.100080\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAderoyeje TG, Erhuanga OG (2024) A Review on the Exploration of Genomic Approaches to Malaria Prevention and Treatment in Nigeria, \u003cem\u003eJ. Appl. Sci. Environ. Manag.\u003c/em\u003e, vol. 28, no. 9, pp. 2937\u0026ndash;2941, Sep. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4314/jasem.v28i9.40\u003c/span\u003e\u003cspan address=\"10.4314/jasem.v28i9.40\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eManning L, Davis TME (2013) The Mechanistic, Diagnostic and Prognostic Utility of Biomarkers in Severe Malaria, \u003cem\u003eBiomark. Med.\u003c/em\u003e, vol. 7, no. 3, pp. 363\u0026ndash;380, Jun. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2217/bmm.13.50\u003c/span\u003e\u003cspan address=\"10.2217/bmm.13.50\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFoko LPK et al (2022) The spectrum of clinical biomarkers in severe malaria and new avenues for exploration, \u003cem\u003eVirulence\u003c/em\u003e, vol. 13, no. 1, pp. 634\u0026ndash;653, Dec. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/21505594.2022.2056966\u003c/span\u003e\u003cspan address=\"10.1080/21505594.2022.2056966\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHunter DJ, Losina E, Guermazi A, Burstein D, Lassere MN, Kraus V (May 2010) A Pathway and Approach to Biomarker Validation and Qualification for Osteoarthritis Clinical Trials. Curr Drug Targets 11(5):536\u0026ndash;545\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOgbadoyi E, Tsado R (2009) Renal and Hepatic Dysfunction in Malaria Patients in Minna, North Central Nigeria, Nov. Accessed: Mar. 15, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.semanticscholar.org/paper/Renal-and-Hepatic-Dysfunction-in-Malaria-Patients-Ogbadoyi-Tsado/eb81837108481004f9f296447592af32aaa31726\u003c/span\u003e\u003cspan address=\"https://www.semanticscholar.org/paper/Renal-and-Hepatic-Dysfunction-in-Malaria-Patients-Ogbadoyi-Tsado/eb81837108481004f9f296447592af32aaa31726\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOyewole I, Senusie S, Mansaray M (2010) Plasmodium Falciparum-Induced Kidney and Liver Dysfunction in Malaria Patients in Freetown, Sierra Leone, \u003cem\u003eSierra Leone J. Biomed. Res.\u003c/em\u003e, vol. 2, no. 1, pp. 70\u0026ndash;74, Jul. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4314/sljbr.v2i1.56611\u003c/span\u003e\u003cspan address=\"10.4314/sljbr.v2i1.56611\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh G, Urhekar A, Singh R, Maheshwari UM, Samant P (2015) Alteration in biochemical parameters in malaria patients. Plasmodium falciparum vs. Plasmodium vivax, Accessed: Mar. 15, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.semanticscholar.org/paper/Alteration-in-biochemical-parameters-in-malaria-vs.-Singh-Urhekar/ad6358a5192eb5417f58a2ff2cafdf3eaa7fee3a\u003c/span\u003e\u003cspan address=\"https://www.semanticscholar.org/paper/Alteration-in-biochemical-parameters-in-malaria-vs.-Singh-Urhekar/ad6358a5192eb5417f58a2ff2cafdf3eaa7fee3a\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHashmi F, Aqeel S, Zuberi UF, Khan W (Sep. 2023) A systematic review and meta-analysis of inflammatory biomarkers associated with malaria infection and disease severity. Cytokine 169:156305. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.cyto.2023.156305\u003c/span\u003e\u003cspan address=\"10.1016/j.cyto.2023.156305\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBiemba G, Gordeuk VR, Thuma PE, Mabeza GF, Weiss G (Jan. 1998) Prolonged macrophage activation and persistent anaemia in children with complicated malaria. Trop Med Int Health 3(1):60\u0026ndash;65. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1046/j.1365-3156.1998.00168.x\u003c/span\u003e\u003cspan address=\"10.1046/j.1365-3156.1998.00168.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAwandare GA et al (2006) Increased Levels of Inflammatory Mediators in Children with Severe \u003cem\u003ePlasmodium falciparum\u003c/em\u003e Malaria with Respiratory Distress, \u003cem\u003eJ. Infect. Dis.\u003c/em\u003e, vol. 194, no. 10, pp. 1438\u0026ndash;1446, Nov. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1086/508547\u003c/span\u003e\u003cspan address=\"10.1086/508547\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGil-G\u0026oacute;mez R, Blasco-Alonso J, S\u0026aacute;nchez-Y\u0026aacute;\u0026ntilde;ez P, Rosa-Camacho V, Manso GM (2017) Niveles de neopterina y s\u0026iacute;ndrome de respuesta inflamatoria sist\u0026eacute;mica en pacientes cr\u0026iacute;ticos pedi\u0026aacute;tricos, \u003cem\u003eAn. Pediatr\u0026iacute;a\u003c/em\u003e, vol. 87, no. 6, pp. 343\u0026ndash;349, Dec. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.anpedi.2016.11.011\u003c/span\u003e\u003cspan address=\"10.1016/j.anpedi.2016.11.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlankson SO et al (Jan. 2023) High Plasma Levels of Neopterin Are Associated with Increased Mortality among Children with Severe Malaria in Benin. Diagnostics 13(3):528. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/diagnostics13030528\u003c/span\u003e\u003cspan address=\"10.3390/diagnostics13030528\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArya A, Kojom Foko LP, Chaudhry S, Sharma A, Singh V (2021) Artemisinin-based combination therapy (ACT) and drug resistance molecular markers: A systematic review of clinical studies from two malaria endemic regions \u0026ndash; India and sub-Saharan Africa, \u003cem\u003eInt. J. Parasitol. Drugs Drug Resist.\u003c/em\u003e, vol. 15, pp. 43\u0026ndash;56, Apr. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ijpddr.2020.11.006\u003c/span\u003e\u003cspan address=\"10.1016/j.ijpddr.2020.11.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarwa K et al (2022) Therapeutic efficacy of artemether-lumefantrine, artesunate-amodiaquine and dihydroartemisinin-piperaquine in the treatment of uncomplicated Plasmodium falciparum malaria in Sub-Saharan Africa: A systematic review and meta-analysis, \u003cem\u003ePLOS ONE\u003c/em\u003e, vol. 17, no. 3, p. e0264339, Mar. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0264339\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0264339\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUrsos LMB, Roepe PD (2002) Chloroquine resistance in the malarial parasite, \u003cem\u003ePlasmodium falciparum\u003c/em\u003e, \u003cem\u003eMed. Res. Rev.\u003c/em\u003e, vol. 22, no. 5, pp. 465\u0026ndash;491, Sep. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/med.10016\u003c/span\u003e\u003cspan address=\"10.1002/med.10016\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaird JK (Nov. 2004) Chloroquine Resistance in \u003cem\u003ePlasmodium vivax\u003c/em\u003e. Antimicrob Agents Chemother 48(11):4075\u0026ndash;4083. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/AAC.48.11.4075-4083.2004\u003c/span\u003e\u003cspan address=\"10.1128/AAC.48.11.4075-4083.2004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChinappi M, Via A, Marcatili P, Tramontano A (Nov. 2010) On the Mechanism of Chloroquine Resistance in Plasmodium falciparum. PLoS ONE 5(11):e14064. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0014064\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0014064\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAshley EA et al (2014) Spread of Artemisinin Resistance in \u003cem\u003ePlasmodium falciparum\u003c/em\u003e Malaria, \u003cem\u003eN. Engl. J. Med.\u003c/em\u003e, vol. 371, no. 5, pp. 411\u0026ndash;423, Jul. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1056/NEJMoa1314981\u003c/span\u003e\u003cspan address=\"10.1056/NEJMoa1314981\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlasco B, Leroy D, Fidock DA (Aug. 2017) Antimalarial drug resistance: linking Plasmodium falciparum parasite biology to the clinic. Nat Med 23(8):917\u0026ndash;928. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nm.4381\u003c/span\u003e\u003cspan address=\"10.1038/nm.4381\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO WHO: World malaria report, 2013 - Google Scholar. Accessed: Oct. 15, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://scholar.google.com/scholar_lookup?title=World%20Malaria%20Report%202019\u0026amp;publication_year=2019\u0026amp;author=WHO\u003c/span\u003e\u003cspan address=\"https://scholar.google.com/scholar_lookup?title=World%20Malaria%20Report%202019\u0026amp;publication_year=2019\u0026amp;author=WHO\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eByakika-Kibwika P, Lamorde M, Mayanja-Kizza H, Khoo S, Merry C, Van Geertruyden J-P (2011) Artemether-Lumefantrine Combination Therapy for Treatment of Uncomplicated Malaria: The Potential for Complex Interactions with Antiretroviral Drugs in HIV-Infected Individuals, \u003cem\u003eMalar. Res. Treat.\u003c/em\u003e, vol. pp. 1\u0026ndash;5, Apr. 2011. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4061/2011/703730\u003c/span\u003e\u003cspan address=\"10.4061/2011/703730\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZwang J et al (Dec. 2012) Comparing changes in haematologic parameters occurring in patients included in randomized controlled trials of artesunate-amodiaquine vs single and combination treatments of uncomplicated falciparum in sub-Saharan Africa. Malar J 11(1):25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2875-11-25\u003c/span\u003e\u003cspan address=\"10.1186/1475-2875-11-25\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchramm B et al (Dec. 2013) Efficacy of artesunate-amodiaquine and artemether-lumefantrine fixed-dose combinations for the treatment of uncomplicated Plasmodium falciparum malaria among children aged six to 59 months in Nimba County, Liberia: an open-label randomized non-inferiority trial. Malar J 12(1):251. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2875-12-251\u003c/span\u003e\u003cspan address=\"10.1186/1475-2875-12-251\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWMA, (2000) World Medical Association Declaration of Helsinki: Ethical Principles for Medical Research Involving Human Subjects Revised October 7, 2000, \u003cem\u003eHIV Clin. Trials\u003c/em\u003e, vol. 2, no. 1, pp. 92\u0026ndash;95, Feb. 2001. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1310/GTFR-2DRX-M6YE-ELXR\u003c/span\u003e\u003cspan address=\"10.1310/GTFR-2DRX-M6YE-ELXR\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRawlings A, Ehiorobo J, Izinyon C (2020) Assessment of Rainfall Seasonality and Distribution Pattern in the South-South Region of Nigeria, vol. 17, pp. 4040\u0026ndash;4059, Apr\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIbebuchi CC, Abu I-O (May 2023) Rainfall variability patterns in Nigeria during the rainy season. Sci Rep 13(1):7888. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-023-34970-7\u003c/span\u003e\u003cspan address=\"10.1038/s41598-023-34970-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlugbenga MA, Obot O-OA, Oluwatoyin HA (2018) Effect of Artemisinin Based Combination Therapy on the Liver and Kidney of Patient Attending University Health Centre. Niger J Pharm Appl Sci Res 7(2):28\u0026ndash;32\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchumann G, Klauke R (Jan. 2003) New IFCC reference procedures for the determination of catalytic activity concentrations of five enzymes in serum: preliminary upper reference limits obtained in hospitalized subjects. Clin Chim Acta Int J Clin Chem 327:1\u0026ndash;2. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/s0009-8981(02)00341-8\u003c/span\u003e\u003cspan address=\"10.1016/s0009-8981(02)00341-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCeriotti F et al (2010) Common reference intervals for aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) in serum: results from an IFCC multicenter study, \u003cem\u003eClin. Chem. Lab. Med.\u003c/em\u003e, vol. 48, no. 11, pp. 1593\u0026ndash;1601, Nov. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1515/CCLM.2010.315\u003c/span\u003e\u003cspan address=\"10.1515/CCLM.2010.315\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAgarwal AK, Katiyar GD, Khan S, Chaudhary BC, Singh A, Sharma M (May 2020) Evaluation of liver function test in malaria positive cases in tertiary care hospital of Bareilly. Int J Community Med Public Health 7(6):2193\u0026ndash;2202. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.18203/2394-6040.ijcmph20202470\u003c/span\u003e\u003cspan address=\"10.18203/2394-6040.ijcmph20202470\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMišić IDR, Mitić SS, Kostić DA, Tošić SB, Pecev-Marinković ET, Miletić AS (2021) Kinetic-spectrophotometric approach to the modified Berthelot procedure for serum urea determination, \u003cem\u003eChem. Pap.\u003c/em\u003e, vol. 75, no. 2, pp. 565\u0026ndash;574, Feb. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11696-020-01315-x\u003c/span\u003e\u003cspan address=\"10.1007/s11696-020-01315-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarakala V, Arnadi Ramachandrayya ASSS, Mala M, Kumar A (Oct. 2012) SERUM CREATININE ASSAY: ENZYMATIC VS KINETIC JAFFE\u0026rsquo;S METHOD. J Evol Med Dent Sci 1:258\u0026ndash;264. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.14260/jemds/54\u003c/span\u003e\u003cspan address=\"10.14260/jemds/54\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVani K, Bodepudi SR, Praveen V Study of Creatinine Estimation by Enzymatic and Kinetic Jaffe\u0026rsquo;s Method\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWhite NJ (2004) Antimalarial drug resistance, \u003cem\u003eJ. Clin. Invest.\u003c/em\u003e, vol. 113, no. 8, pp. 1084\u0026ndash;1092, Apr. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1172/JCI21682\u003c/span\u003e\u003cspan address=\"10.1172/JCI21682\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbebe W, Woldesenbet D (Aug. 2025) A review on molecular markers of Plasmodium falciparum. J Parasit Dis. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s12639-025-01840-0\u003c/span\u003e\u003cspan address=\"10.1007/s12639-025-01840-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVasquez M, Zuniga M, Rodriguez A (Nov. 2021) Oxidative Stress and Pathogenesis in Malaria. Front Cell Infect Microbiol 11:768182. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fcimb.2021.768182\u003c/span\u003e\u003cspan address=\"10.3389/fcimb.2021.768182\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChandrashekhar VN et al (2022) Impact of oxidative stress in response to malarial infection during pregnancy: Complications, histological changes, and pregnancy outcomes, \u003cem\u003eTrop. Parasitol.\u003c/em\u003e, vol. 12, no. 1, p. 21, Jun. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4103/tp.TP_18_20\u003c/span\u003e\u003cspan address=\"10.4103/tp.TP_18_20\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh G, URHEKAR A, Singh R, MAHESHWARI U, SAMANT P, Al A (2015) Alteration in biochemical parameters in malaria patients. Plasmodium falciparum vs. Plasmodium vivax, \u003cem\u003eJ. Microbiol. Antimicrob. AGENTS\u003c/em\u003e, vol. 1, Jan\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKochar DK et al (Jul. 2003) Hepatocyte dysfunction and hepatic encephalopathy in Plasmodium falciparum malaria. QJM 96(7):505\u0026ndash;512. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/qjmed/hcg091\u003c/span\u003e\u003cspan address=\"10.1093/qjmed/hcg091\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlusola AO (2018) Effect of Plasmodium Falciparum on Liver Function Parameters of Children in Akoko Area of Ondo State, Nigeria. J Diagn 5(1):1\u0026ndash;5. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.18488/journal.98.2018.51.1.5\u003c/span\u003e\u003cspan address=\"10.18488/journal.98.2018.51.1.5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReuling IJ et al (Oct. 2018) Liver Injury in Uncomplicated Malaria is an Overlooked Phenomenon: An Observational Study. eBioMedicine 36:131\u0026ndash;139. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ebiom.2018.09.018\u003c/span\u003e\u003cspan address=\"10.1016/j.ebiom.2018.09.018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTangpukdee N et al (2006) Minor liver profile dysfunctions in Plasmodium vivax, P. malariae and P. ovale patients and normalization after treatment. Korean J Parasitol 44(4):295. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3347/kjp.2006.44.4.295\u003c/span\u003e\u003cspan address=\"10.3347/kjp.2006.44.4.295\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOdedra A, Woodford J, Chalon S, Barber BE, McCarthy JS (2022) Posttreatment Transaminase Elevations in Controlled Human Malaria Infection and Naturally Acquired Malaria, \u003cem\u003eJ. Infect. Dis.\u003c/em\u003e, vol. 225, no. 5, pp. 926\u0026ndash;927, Mar. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/infdis/jiab502\u003c/span\u003e\u003cspan address=\"10.1093/infdis/jiab502\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCheaveau J et al (Jun. 2019) The Impact of Malaria on Liver Enzymes: A Retrospective Cohort Study (2010\u0026ndash;2017). Open Forum Infect Dis 6(6):ofz234. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/ofid/ofz234\u003c/span\u003e\u003cspan address=\"10.1093/ofid/ofz234\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTrang TTM et al (Nov. 1992) Acute Renal Failure in Patients with Severe Falciparum Malaria. Clin Infect Dis 15(5):874\u0026ndash;880. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/clind/15.5.874\u003c/span\u003e\u003cspan address=\"10.1093/clind/15.5.874\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMishra SK, Das BS (Jul. 2008) Malaria and Acute Kidney Injury. Semin Nephrol 28(4):395\u0026ndash;408. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.semnephrol.2008.04.007\u003c/span\u003e\u003cspan address=\"10.1016/j.semnephrol.2008.04.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eConroy AL et al (Apr. 2016) Acute Kidney Injury Is Common in Pediatric Severe Malaria and Is Associated With Increased Mortality. Open Forum Infect Dis 3(2):ofw046. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/ofid/ofw046\u003c/span\u003e\u003cspan address=\"10.1093/ofid/ofw046\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAfolayan FM et al (Nov. 2020) Acute Kidney Injuries in Children with Severe Malaria: A comparative study of diagnostic criteria based on serum cystatin C and creatinine levels. Sultan Qaboos Univ Med J 20(4):e312\u0026ndash;e317. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.18295/squmj.2020.20.04.006\u003c/span\u003e\u003cspan address=\"10.18295/squmj.2020.20.04.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatsoulis O, Georgiadou A, Cunnington AJ (Apr. 2021) Immunopathology of Acute Kidney Injury in Severe Malaria. Front Immunol 12:651739. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2021.651739\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2021.651739\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarrington L et al (Dec. 2017) Both inflammatory and regulatory cytokine responses to malaria are blunted with increasing age in highly exposed children. Malar J 16(1):499. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s12936-017-2148-6\u003c/span\u003e\u003cspan address=\"10.1186/s12936-017-2148-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMackintosh CL, Beeson JG, Marsh K (Dec. 2004) Clinical features and pathogenesis of severe malaria. Trends Parasitol 20(12):597\u0026ndash;603. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.pt.2004.09.006\u003c/span\u003e\u003cspan address=\"10.1016/j.pt.2004.09.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWilairatana P et al (Nov. 2021) C-reactive protein as an early biomarker for malaria infection and monitoring of malaria severity: a meta-analysis. Sci Rep 11(1):22033. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-021-01556-0\u003c/span\u003e\u003cspan address=\"10.1038/s41598-021-01556-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBhardwaj N, Ahmed M, Sharma S, Nayak A, Anvikar A, Pande V (2019) C-reactive protein as a prognostic marker of Plasmodiumfalciparum malaria severity. J Vector Borne Dis 56(2):122. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4103/0972-9062.263727\u003c/span\u003e\u003cspan address=\"10.4103/0972-9062.263727\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown AE, Webster HK, Teja-Isavadharm P, Keeratithakul D (Oct. 1990) Macrophage activation in falciparum malaria as measured by neopterin and interferon-gamma. Clin Exp Immunol 82(1):97\u0026ndash;101. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1365-2249.1990.tb05410.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1365-2249.1990.tb05410.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTe Witt R et al (Dec. 2010) Neopterin and procalcitonin are suitable biomarkers for exclusion of severe Plasmodium falciparum disease at the initial clinical assessment of travellers with imported malaria. Malar J 9(1):255. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2875-9-255\u003c/span\u003e\u003cspan address=\"10.1186/1475-2875-9-255\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNagaraj N, Berwal PK, Srinivas A, Prakash P, Ramesh MS, Berwal A (2018) Correlation of hepatorenal dysfunction in pediatric malaria. Trop Parasitol 8(2):83\u0026ndash;87. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4103/tp.TP_2_16\u003c/span\u003e\u003cspan address=\"10.4103/tp.TP_2_16\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePlewes K et al (Mar. 2014) Correlation of biomarkers for parasite burden and immune activation with acute kidney injury in severe falciparum malaria. Malar J 13:91. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2875-13-91\u003c/span\u003e\u003cspan address=\"10.1186/1475-2875-13-91\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOjo RJ, Jonathan IG, Adams MD, Gyebi G, Longdet IY (Oct. 2022) Renal and hepatic dysfunction parameters correlate positively with gender among patients with recurrent malaria cases in Birnin Kebbi, Northwest Nigeria. Egypt J Intern Med 34(1):77. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s43162-022-00164-2\u003c/span\u003e\u003cspan address=\"10.1186/s43162-022-00164-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli MO, Tamomh AG, Abdulrahman NA, Ibrahim NS, Eltayeb MA, Osman FI (Sep. 2025) Exploring the Effect of Malaria on Biochemical Parameters Among Sudanese Patients with Complicated Falciparum Malaria. Clin Lab 71(9). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.7754/Clin.Lab.2025.250208\u003c/span\u003e\u003cspan address=\"10.7754/Clin.Lab.2025.250208\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan JM, Hindoliya BS, Paul A, Shahrukh M (2023) A Comparative Study Of Liver Enzymes In The Individuals With Malarial Infection And Normal Healthy Individuals, \u003cem\u003eEurasian J. Anal. Chem.\u003c/em\u003e, vol. 18, no. 1, pp. 12\u0026ndash;16, Dec\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGodse RR (2013) Hematological and biochemical evaluation in malaria patients with clinical correlation, Accessed: Nov. 22, 2025. [Online]. Available: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.semanticscholar.org/paper/Hematological-and-biochemical-evaluation-in-malaria-Rr/13e5817eec55076385af5261e5ea023f01434d00\u003c/span\u003e\u003cspan address=\"https://www.semanticscholar.org/paper/Hematological-and-biochemical-evaluation-in-malaria-Rr/13e5817eec55076385af5261e5ea023f01434d00\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLockard RD, Wilson ME, Rodr\u0026iacute;guez NE (2019) Sex-Related Differences in Immune Response and Symptomatic Manifestations to Infection with \u003cem\u003eLeishmania\u003c/em\u003e Species, \u003cem\u003eJ. Immunol. Res.\u003c/em\u003e, vol. pp. 1\u0026ndash;14, Jan. 2019. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1155/2019/4103819\u003c/span\u003e\u003cspan address=\"10.1155/2019/4103819\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRuggieri A, Gagliardi MC, Anticoli S (Oct. 2018) Sex-Dependent Outcome of Hepatitis B and C Viruses Infections: Synergy of Sex Hormones and Immune Responses? Front Immunol 9:2302. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2018.02302\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2018.02302\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown R, Goulder P, Matthews PC (2022) Sexual Dimorphism in Chronic Hepatitis B Virus (HBV) Infection: Evidence to Inform Elimination Efforts. Wellcome Open Res 7:32. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.12688/wellcomeopenres.17601.3\u003c/span\u003e\u003cspan address=\"10.12688/wellcomeopenres.17601.3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDuncombe CJ et al Sex-Based Differences to Plasmodium Infection in the Placebo Arms of Controlled Human Malaria Infection (CHMI) Trials in Malaria-Na\u0026iuml;ve Populations: A Pooled Analysis, Jan. 13, 2025, \u003cem\u003eSocial Science Research Network, Rochester, NY\u003c/em\u003e: 5087394. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2139/ssrn.5087394\u003c/span\u003e\u003cspan address=\"10.2139/ssrn.5087394\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoon JJ, Cho S-Y (2001) Incidence patterns of vivax malaria in civilians residing in a high-risk county of Kyonggi-do (Province), Republic of Korea, \u003cem\u003eKorean J. Parasitol.\u003c/em\u003e, vol. 39, no. 4, pp. 293\u0026ndash;299, Dec. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3347/kjp.2001.39.4.293\u003c/span\u003e\u003cspan address=\"10.3347/kjp.2001.39.4.293\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePathak S et al (2012) Age-Dependent Sex Bias in Clinical Malarial Disease in Hypoendemic Regions, \u003cem\u003ePLOS ONE\u003c/em\u003e, vol. 7, no. 4, p. e35592, Apr. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0035592\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0035592\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBriggs J et al (2020) Sex-based differences in clearance of chronic Plasmodium falciparum infection, \u003cem\u003eeLife\u003c/em\u003e, vol. 9, p. e59872, Oct. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.7554/eLife.59872\u003c/span\u003e\u003cspan address=\"10.7554/eLife.59872\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"the-egyptian-journal-of-internal-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [The Egyptian Journal of Internal Medicine](https://ejim.springeropen.com/)","snPcode":"43162","submissionUrl":"https://submission.springernature.com/new-submission/43162/3","title":"The Egyptian Journal of Internal Medicine","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"malaria, ALT, AST, urea, creatinine, CRP, neopterin, artemether, lumefantrine","lastPublishedDoi":"10.21203/rs.3.rs-8673475/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8673475/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eMalaria remains a major health concern in many parts of the world. Globally, about 260\u0026nbsp;million malaria cases and 600,000 malaria-related deaths, caused by different \u003cem\u003ePlasmodium spp\u003c/em\u003e., are recorded annually. Biochemical markers play a crucial role in the diagnosis or prognosis of malaria, and the blood levels of the associated biomarkers could be altered upon infection or post-treatment with antimalarials. This study aimed to investigate the impact of artemether-lumefantrine combination therapy on the serum levels of biochemical markers in malaria patients.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003eBlood samples were collected from 30 malaria patients and matched with those from 20 healthy control subjects. Parasitaemia and blood levels of selected liver function, kidney function, and inflammation markers were measured in malaria patients and healthy control subjects. The malaria patients were treated with the standard dose of artemether-lumefantrine combination therapy. After recovery from malaria, parasitaemia and blood levels of the selected markers were measured again.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eDuring infection with \u003cem\u003eP. falciparum\u003c/em\u003e, serum ALT and AST were reduced, parasitaemia and serum CRP were increased, while serum urea, creatinine, and neopterin remained statistically unchanged when compared with their levels in healthy control subjects. After treatment with artemether-lumefantrine combination therapy, serum creatinine and CRP levels were reduced, AST was increased, while parasitaemia, ALT, urea, and neopterin levels remained statistically unchanged when compared with their levels in malaria patients pre-treatment.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eTaken together, our data demonstrated that infection with \u003cem\u003eP. falciparum\u003c/em\u003e can modulate the serum levels of key hepatic, renal, and inflammatory markers. Furthermore, treatment with artemether-lumefantrine combination therapy is effective in reversing some of the pathological changes caused by \u003cem\u003eP. falciparum\u003c/em\u003e in malaria patients.\u003c/p\u003e","manuscriptTitle":"Comparative analysis of biochemical parameters in the blood of malaria patients: correlation with treatment response to artemether-lumefantrine combination therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-11 05:24:26","doi":"10.21203/rs.3.rs-8673475/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2026-02-15T18:03:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-14T00:19:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"291778546360803904078655053521408775106","date":"2026-02-14T00:15:23+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-06T19:59:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"143559848696398308304199029109716568442","date":"2026-02-06T15:32:41+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"70663990994060276474831801380210692792","date":"2026-02-06T03:56:10+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-05T20:17:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-04T08:52:16+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-29T05:58:35+00:00","index":"","fulltext":""},{"type":"submitted","content":"The Egyptian Journal of Internal Medicine","date":"2026-01-22T22:32:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"the-egyptian-journal-of-internal-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [The Egyptian Journal of Internal Medicine](https://ejim.springeropen.com/)","snPcode":"43162","submissionUrl":"https://submission.springernature.com/new-submission/43162/3","title":"The Egyptian Journal of Internal Medicine","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ce6159bd-1edb-4258-a0c4-517d5c4c67e0","owner":[],"postedDate":"February 11th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-02-11T05:24:26+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-11 05:24:26","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8673475","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8673475","identity":"rs-8673475","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}