Prevalence of Helicobacter pylori among Sudanese patients diagnosed with colon polyps and colon cancer using immunohistochemistry technique.

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Aziza K. Mohamed, Nazik M. Elhassan, Zahra A. Awhag, Fadwa S. Ali, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-31738/v3 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Jul, 2020 Read the published version in BMC Research Notes → Version 3 posted 2 You are reading this latest preprint version Show more versions Abstract Objectives: Infection with the bacteria Helicobacter pylori has been classified as class one carcinogen associated with increasing susceptibility of gastritis and gastric carcinoma. This study is aiming at investigating the prevalence of H. pylori among colon polyps and colon cancer patients. A descriptive cross-sectional hospital-based study was conducted between February and June 2017. Sixty-nine formalin-fixed paraffin blocks collected from colon polyps and colon cancer patients to detect H. pylori using immunohistochemistry technique. Results: Of the Sixty-nine patients included in the study, 39 (56.5%) males and 30 (43.5%) were females, their age ranged from 21 to 80 years with a mean age of 47.1±19.7. Of the 69 colon polyps and colon cancer patients, 44 (63.8%) were diagnosed as adenocarcinoma, 10 (14.5%) colitis, 15 (21.7%) juvenile polyposis syndrome. The results of immunohistochemistry technique showed the presence of 16 (23.2%) positive patients for H. pylori infection. Of these 16, 13 (81.3%) patients were diagnosed with adenocarcinoma and 3 (18.7%) patients were diagnosed with juvenile polyps. The results of H. pylori detection among the different colon polyps and colon cancer patients were showing a statistically significant association for H. pylori infection and adenocarcinoma, P value 0.028. Cancer Biology Colon polyps Colon cancer Helicobacter pylori Immunohistochemistry Figures Figure 1 Introduction Colorectal cancer (CRC) is accounted as the third most common malignancy and the third most common cause of death due to cancer in both men and women in the US [1]. CRC mostly arises from adenomatous polyps (adenomas) and from hyperplastic polyps [2, 3]. Early diagnosis and surgical removal of these polyps have associated with the decreased in the incidence of mortality [4-6]. Therefore, clinicians started to focus in recent years on the prevention measures that decrease the incidence of developing CRC; although, researchers started to explore the role of several infectious organism and their ability to increase or induce CRC [7-10]. For instance, many epidemiological studies have linked the infection of Helicobacter pylori to colorectal neoplasm either through high prevalence of H. pylori seropositivity among CRC or colorectal polyp patients [11-14], or through the presence of bacterial products and their trophic effects on colon mucosa [15-18]. Moreover, few studies have linked the presence of H. pylori in the stomach or colon with colon cancer and/or polyps [19-24]. It is well known that H. pylori predisposes to develop gastric cancer precursor lesions, thus it has been classified as class 1 carcinogen [25]. A recent meta-analysis correlating between H. pylori and extra-gastric malignancies revealed a statistically significant relationship of H. pylori infection with both colon cancer and polyps [26]. Also, H. pylori infection linked with colorectal lesions appeared to be more common in African Americans compared to the Caucasian population in the US [27, 28]. Epidemiological studies have confirmed a causal relationship between H. pylori and gastric cancer, and the colonic phenotype of H. pylori -related intestinal metaplasia (IM) [29]. Thus, association of H. pylori in various gastrointestinal cancers has been investigated, moreover, Helicobacter DNAs were prevalent in more than 50 percent of hepatobiliary cancer cases [30]. Helicobacter species, which may colonize the biliary tract, have been implicated as a possible cause of hepatobiliary diseases ranging from chronic cholecystitis and primary sclerosing cholangitis to gall-bladder carcinoma and primary hepatic carcinomas [31]. Therefore, the hypothesis that H. pylori would also be associated with colon lesions needs to be investigated. In Sudan, no reports addressing this manner were existed. Previous studies investigating the seroprevalence of H. pylori among Sudanese using ELISA and rapid immunochromatographic tests for the detection of H. pylori IgM and IgG antibodies, reported a seroprevalence ranging from 20% up to 70% [32-35]. Therefore, the aim of this study was to investigate the presence of H. pylori infections among Sudanese patients diagnosed with colon polyps and colon cancer and to correlate between its presence and the type of the lesions. Materials And Methods Sample and data collection This is a preliminary, descriptive study aimed to investigate the frequency of H. pylori infections among Sudanese patients diagnosed with colon cancer. Data were collected from 69 patients attended the National Laboratory and Alrahma Laboratory between February-June 2017. During colonoscopy, presence of abnormal tissue, such as clumps of cells; polyps, formed on the inside of the colon, the pathologists obtained colon biopsies. Colon biopsies were fixed with formalin and processed into paraffin embedded blocks. Formalin-fixed paraffin blocks were used for the immunohistochemical detection of H. pylori . Ethical approval was previously obtained by the pathologists of each hospital before colon polyps’ biopsies were taken. Preparation of the formalin fixed paraffin blocks Four sections from each formalin-fixed paraffin block to increase detection sensitivity were obtained with a thickness of 4µm using Rotary microtome (LEICA RM2125RT). All sections were de-waxed with two changes of Xylene for 3 minutes and then dehydrated in descending concentrations of Methanol starting from absolute Methanol through 90% and lastly, a concentration of 70% for 2 minutes in each concentration, and then washed using distilled water. Immunohistochemistry diagnosis Immunohistochemistry diagnosis was performed on all the obtained sections. Known gastric sections containing H. pylori infection was used as positive and negative controls; for the negative control the primary antibody incubation step was omitted. All sections were pretreated to retrieve antigens at 97 o C for 10 minutes in citrate buffer solution, and then sections were blocked by 3% Hydrogen peroxide and absolute Methanol for 20 minutes at humidified chamber. Afterward, sections were blocked into Bovine serum Albumin (Thermo Fisher Scientific, Germany). A rabbit polyclonal antibody ULC3R (BioGenex, USA) (prepared from tissue culture supernatant diluted in PBS, pH 7.6 containing 5% BSA and 0.09% Sodium Azaide) against H. pylori was applied for 40 minutes, then washed in buffer solution for 5 minutes. Then, a polymer solution was applied for 15 minutes, and washed in buffer solution for 5 minutes. Chromogen solution was added for 10 minutes, and washed in distilled water. Finally, Mayer’s Hematoxylin was added for 2 minutes, and then, sections were blued using running distilled water for 5 minutes. After bluing, sections were dehydrated, cleared, and mounted in DPX. Prepared sections were investigated microscopically by two experts’ pathologists blindly without knowing the duplication of slides sections of each patient using X40 lens. Results were recorded into categories of positive and negative results; the dot like shape denoted the coccoid form of the organism as describe previously [36-39]. Statistical analysis: Descriptive data were analyzed using the Statistical Package for Social Science (SPSS-v20). Pearson Chi-square test was used to test the association of H. pylori infection with the different types of lesions. A P value < 0.05 was considered as a statistically significant. Results Of the 69 patients, there were 30 (43.5%) females and 39 (56.5%) males, their ages ranged from 21 to 80 years with a mean age 47.1±19.8. 44 (63.8%) patients were diagnosed with adenocarcinoma, 10 (14.5%) patients were colitis, and 15 (21.7%) patients were juvenile polyposis syndrome. No statistically significant was observed for the association of gender and the pathological condition of each patient, P value = 0.649. Out of the 69 patients, 16 (23.2%) patients were positive for H. pylori infection. These 16 positive patients included; 13 (81.3%) patients diagnosed with adenocarcinoma and 3 (18.7%) patients diagnosed with juvenile polyps. The correlation between presence of H. pylori infection and the histopathological condition of patients were positively correlated (P value 0.028) (Table 1). In respect to Immunohistochemistry diagnosis, the bacteria were prominent and easier to detect in the immune-stained sections in several patterns including organisms attached to the epithelial cells or within the superficial mucus. And in some cases, the bacteria were masked by inspissated mucus or being positioned flat and closely opposed to the epithelial surface. Regarding the morphological appearance of the organism; H. pylori stained brown in color and take a dot and small curved shape in different sizes (Figure 1). Discussion The exact role of H. pylori in the induction of colon cancer is still a debate between the scientific researcher communities; this is attributed to the controversial results obtained. In previous studies, H. pylori were linked to the development of gastric cancer [40], while others reported paradoxical results showing no association between H. pylori and gastric cancer susceptibility [41-43]. However, reports from Sudan regarding the possible link between H. pylori and colon cancer are scarce. Therefore, in the present study, we examined the presence of H. pylori using immunohistochemistry technique on colon polyps and colon cancer lesions of Sudanese patients underwent colonoscopy. The results obtained from this study showed a positive correlation between the presence of H. pylori infection and the histopathological diagnosis, as H. pylori was prevalent in higher frequency in patients diagnosed with adenocarcinoma compared to those diagnosed as juvenile polyposis syndrome, and the result was statistically significant. This result also agrees with studies conducted by Jones et al, and Grahn et al., they investigated the presence of H. pylori among 59 patients diagnosed with colon adenocarcinomas using immunohistochemistry technique, and 77 colon and rectum cancer patients using molecular technique, correspondingly [44, 45]. Jones et al., reported that H. pylori were detected in 10/59 adenocarcinoma cases which represent about 16.9% of the total cancer cases studied [44]. While, Grahn et al., showed that H. pylori were present in 27% of the patients; among the studied colon cancer, H. pylori were present in 11/42 (26%) patients [45]. Although, several studies failed to demonstrate any association between H. pylori and colon cancer, or even if this microorganism can colonize the colon [46-50]. This could only be attributed to the ability of demonstrating the H. pylori bacteria, which was achieved by the aid of the immunohistochemistry technique that allowed a better localization of H. pylori within the various colon lesions of colitis, polyps, and adenocarcinoma included in this study. Interestingly, several theories were proposed regarding the exact role by which H. pylori induced colon cancer, one hypothesis is that colon cancer can be induced by toxins produced by H. pylori ; however, this theory was based only on serological data [43, 51-53]. Furthermore, some studies showed that colitis and colon cancer were also developed in experimental mice models infected with H. hepaticus [54]. Accordingly, the development of colon cancer seems most likely due to the interaction between toxins produced by the bacteria and the immune cells of the mice [54]. Therefore, the results we obtained from our study showing that H. pylori were present, nevertheless, it means that H. pylori infection is responsible for the induction and the development of colon cancer, since the presence of H. pylori could be encountered as post-cancer incidence. This however, still requires more complicated experimental studies to investigate this hypothesis; yet, the role of H. pylori cannot be excluded due to this hypothesis. Therefore, this preliminary report needs further advanced experimental investigations to enable the determination on the exact mechanisms by which H. pylori can induce colon cancer. Conclusion This study was able to demonstrate the presence of H. pylori in colon polyps and colon cancer using immunohistochemistry marker, besides the significant association in the presence of H. pylori with colon adenocarcinoma, indeed further studies are required to elaborate more in-depth about the exact role of H. pylori in the development of colon cancer. Limitations In this study the sample size studied was relatively small, but still a significant association was observed. A bigger sample size of colon cancer lesions and benign colon lesions; non cancer lesions, should be included in future studies to determine the significant association of H. pylori with adenocarcinoma among the Sudanese patients diagnosed with colon cancer. Abbreviations DNA: Deoxyribonucleic acid; CRC: Colorectal cancer; IM: Intestinal Metaplasia. Declarations Ethics approval and consent to participate The study ethical clearance was obtained from University of Khartoum, Faculty of Medical Laboratory Sciences ethical review board. Informed consent was obtained from each participant prior to enrollment using writing and verbal informed consent. Consent to publish Not Applicable. Availability of Data and Materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing interests The authors declare that they have no competing interest s. Funding Not Applicable. Authors' contributions AKM, NME, ZAA, FSA, EES and AMME provided conceptual framework for the project. AKM, NME, ZAA, FSA, ETA, NAM, RH, HAO and ESA participated in the preparation of the samples and performed the diagnosis. NSM, AF, EES, MSM, AA, and AMME performed the data analysis and guidance for data interpretation. AA, MSM, ETA, NAM, RH, ESA, AF, and NSM drafted the manuscript. All authors read and approved the final manuscript. Acknowledgements The authors are grateful to the patients and the staff at the National Laboratory and Al-Rahma Laboratory for their continued collaboration and generous hospitality. Author’s Information Aziza K. Mohamed 1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Nazik M. Elhassan 1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Zahra A. Awhag 1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Fadwa S. Ali 1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Eman T. Ali 1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. 2 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, National University, Sudan, Khartoum, Sudan. Email: [email protected] . Najwa A. Mhmoud 3 Mycetoma research center, Khartoum, Sudan. 4 Department of Microbiology and immunology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Emmanuel E. Siddig 3 Mycetoma research center, University of Khartoum, Khartoum, Sudan. 5 Nile University, School of medicine, Khartoum, Sudan. 6 Department of Histopathology and Cytology, Alfarrabi College for Science and Technology, Khartoum, Sudan. Email: [email protected] . Rowa Hassan 3 Mycetoma Research Center, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Eiman S. Ahmed 3 Mycetoma Research Center, University of Khartoum, Khartoum, Sudan. Email: [email protected] . Azam Fattahi 7 Center for Research and Training in Skin Disease and Leprosy, Tehran University of Medical Sciences, Tehran, Iran. Email: [email protected] . Ayman Ahmed 8 Department of Parasitology and Medical Entomology, Institute of Endemic Diseases, University of Khartoum, Sudan. Email: [email protected] Mohamed S. Muneer 9 Department of Neurology, Mayo Clinic, Jacksonville, FL, USA. 10 Department of Radiology, Mayo Clinic, Jacksonville, FL, USA 11 Department of Internal Medicine, Faculty of Medicine, University of Khartoum, Khartoum, Sudan. Email: [email protected] Hussam A. Osman 12 Biomedical Research Laboratory, Ahfad University for Women, Omdurman, Sudan. Email: [email protected] Nouh S. Mohamed 12 Molecular Biology Department, Biotechnology Lab, Faculty of Pharmacy. Ahfad University for women, Khartoum Sudan. 13 Department of Parasitology and Medical Entomology, Faculty of Medicine, Sinnar University, Sinnar, Sudan. 14 Department of Parasitology and Medical Entomology, Faculty of Medical Laboratory Sciences, Nile University, Khartoum, Sudan. Email: [email protected] . Ali MM. 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Shmuely H, Passaro D, Figer A, Niv Y, Pitlik S, Samra Z, Koren R, Yahav JJTAjog: Relationship between Helicobacter pylori CagA status and colorectal cancer. 2001, 96: 3406. Fireman Z, Trost L, Kopelman Y, Segal A, Sternberg AJTIMAjI: Helicobacter pylori: seroprevalence and colorectal cancer. 2000, 2: 6-9. Hsu W-Y, Lin C-H, Lin C-C, Sung F-C, Hsu C-P, Kao C-H: The relationship between Helicobacter pylori and cancer risk. European journal of internal medicine 2014, 25: 235-240. Bell S, Chisholm S, Owen R, Borriello S, Kamm MJAp, therapeutics: Evaluation of Helicobacter species in inflammatory bowel disease. 2003, 18: 481-486. Table Table 1: Shows the correlation between gender, immunohistochemistry detection of H. pylori with the histopathological diagnosis. Histopathological Diagnosis Total P value Adenocarcinoma Juvenile Polyposis Syndrome Colitis Gender Male 23 (58.9%) 10 (25.6%) 6 (15.5%) 39 (56.5%) 0.649 Female 21 (70.0%) 5 (16.7%) 4 (13.3%) 30 (43.5%) Total 44 (63.8%) 15 (21.7%) 10 (14.5%) 69 (100%) Immunohistochemistry of H. Pylori Negative 31 (58.5%) 12 (22.6%) 10 (18.9%) 53 (76.8%) 0.028 Positive 13 (81.3%) 3 (18.7%) 0 (0.0%) 16 (23.2%) Total 44 (63.8%) 15 (21.7%) 10 (14.5%) 69 (100%) Cite Share Download PDF Status: Published Journal Publication published 06 Jul, 2020 Read the published version in BMC Research Notes → Version 3 posted Submission checks completed at journal 25 Jun, 2020 Editorial decision: Accept 25 Jun, 2020 You are reading this latest preprint version Show more versions 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. 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Siddig","email":"","orcid":"","institution":"University of Khartoum","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Emmanuel","middleName":"E.","lastName":"Siddig","suffix":""},{"id":719466,"identity":"d0b37b60-22b9-4ddf-9e44-69a55a015cf7","order_by":7,"name":"Rowa Hassan","email":"","orcid":"","institution":"University of Khartoum","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Rowa","middleName":"","lastName":"Hassan","suffix":""},{"id":719467,"identity":"1b99b50e-898c-440f-a586-8a0a39064507","order_by":8,"name":"Eiman S. 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Muneer","email":"","orcid":"","institution":"Mayo Clinic's Campus in Florida","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"S.","lastName":"Muneer","suffix":""},{"id":719471,"identity":"3ccef30c-8749-48e1-a6e4-b67e67b81359","order_by":12,"name":"Hussam A. Osman","email":"","orcid":"","institution":"Ahfad University for Women School of Pharmacy","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hussam","middleName":"A.","lastName":"Osman","suffix":""},{"id":719472,"identity":"0d06baa2-dedd-4b6a-a5d7-2026b8b56d6f","order_by":13,"name":"Nouh Saad Mohamed","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzUlEQVRIiWNgGAWjYHACNgh1vAFIFDAwSBCv5cwBBoYDBiRpuZFApBZz9h6zBwx/7OT5bj4+Jv3BwEZOsoH54aMbeLRY9pwxN2BsSzaceTstTeKAQZqxNAObsXEOHi0GN3LMJBgbmBk33AYyDhgcTpzHwMMmTVALw596+w03z5Ckhe1w4oYbPBAtswlqOXOsTIKx7XjyzDNpyRZngH6RbCbkl+PN24AOq7btO3744I2KChs5iePNDx/j0wICzH9QuQSUj4JRMApGwSggDABYl0mEzoxVyQAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0001-6843-3361","institution":"Ahfad University for Women School of Pharmacy","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Nouh","middleName":"Saad","lastName":"Mohamed","suffix":""},{"id":719473,"identity":"19a1ca2e-82b3-4a3f-bc1e-61c75e599bcd","order_by":14,"name":"Ali MM. Edris","email":"","orcid":"","institution":"University of Bisha","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ali","middleName":"MM.","lastName":"Edris","suffix":""}],"badges":[],"createdAt":"2020-05-26 03:48:10","currentVersionCode":3,"declarations":"","doi":"10.21203/rs.3.rs-31738/v3","doiUrl":"https://doi.org/10.21203/rs.3.rs-31738/v3","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s13104-020-05159-2","type":"published","date":"2020-07-06T12:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":1418284,"identity":"d73d2ff6-2c9d-43f0-9b5f-bc72a651250b","added_by":"auto","created_at":"2020-06-25 18:42:25","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":621926,"visible":true,"origin":"","legend":"Detection of H. pylori using IHC staining technique. A: indicates positive control tissue section of known gastric biopsy infected with H. pylori. B, C, and D: indicates tissue sections of patients diagnosed with colon polyps and colon cancer. Black arrows indicate H. pylori stained brown in color and take a dot and small curved shape in different size.","description":"","filename":"fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-31738/v3/fig1.png"},{"id":15667713,"identity":"8f342bcf-2631-427f-a548-611b6c87b8c4","added_by":"auto","created_at":"2021-11-18 13:44:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2445577,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-31738/v3/860805b9-c6fe-41b9-a3f5-53d2ba5b4ec9.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003ePrevalence of \u003cem\u003eHelicobacter pylori\u003c/em\u003e among Sudanese patients diagnosed with colon polyps and colon cancer using immunohistochemistry technique.\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eColorectal cancer (CRC) is accounted as the third most common malignancy and the third most common cause of death due to cancer in both men and women in the US [1]. CRC mostly arises from adenomatous polyps (adenomas) and from hyperplastic polyps [2, 3]. Early diagnosis and surgical removal of these polyps have associated with the decreased in the incidence of mortality [4-6]. Therefore, clinicians started to focus in recent years on the prevention measures that decrease the incidence of developing CRC; although, researchers started to\u0026nbsp; explore the role of several infectious organism and their ability to increase or induce CRC [7-10]. For instance, many epidemiological studies have linked the infection of \u003cem\u003eHelicobacter pylori\u003c/em\u003e to colorectal neoplasm either through high prevalence of \u003cem\u003eH. pylori\u003c/em\u003e seropositivity among CRC or colorectal polyp patients [11-14], or through the presence of bacterial products and their trophic effects on colon mucosa [15-18]. Moreover, few studies have linked the presence of \u003cem\u003eH. pylori\u003c/em\u003e in the stomach or colon with colon cancer and/or polyps [19-24].\u003c/p\u003e\n\u003cp\u003eIt is well known that \u003cem\u003eH. pylori\u003c/em\u003e predisposes to develop gastric cancer precursor lesions, thus it has been classified as class 1 carcinogen [25]. A recent meta-analysis correlating between \u003cem\u003eH. pylori\u003c/em\u003e and extra-gastric malignancies revealed a statistically significant relationship of \u003cem\u003eH. pylori\u003c/em\u003e infection with both colon cancer and polyps [26]. \u0026nbsp;Also, \u003cem\u003eH. pylori\u003c/em\u003e infection linked with colorectal lesions appeared to be more common in African Americans compared to the Caucasian population in the US [27, 28].\u003c/p\u003e\n\u003cp\u003eEpidemiological studies have confirmed a causal relationship between \u003cem\u003eH. pylori\u003c/em\u003e and gastric cancer, and the colonic phenotype of \u003cem\u003eH. pylori\u003c/em\u003e-related intestinal metaplasia (IM) [29]. Thus, association of \u003cem\u003eH. pylori \u003c/em\u003ein various gastrointestinal cancers has been investigated, moreover, \u003cem\u003eHelicobacter\u003c/em\u003e DNAs were prevalent in more than 50 percent of hepatobiliary cancer cases [30]. \u003cem\u003eHelicobacter\u003c/em\u003e species, which may colonize the biliary tract, have been implicated as a possible cause of hepatobiliary diseases ranging from chronic cholecystitis and primary sclerosing cholangitis to gall-bladder carcinoma and primary hepatic carcinomas [31]. Therefore, the hypothesis that \u003cem\u003eH. pylori\u003c/em\u003e would also be associated with colon lesions needs to be investigated. In Sudan, no reports addressing this manner were existed. Previous studies investigating the seroprevalence of \u003cem\u003eH. pylori\u003c/em\u003e among Sudanese using ELISA and rapid immunochromatographic tests for the detection of \u003cem\u003eH. pylori\u003c/em\u003e IgM and IgG antibodies, reported a seroprevalence ranging from 20% up to 70% [32-35]. Therefore, the aim of this study was to investigate the presence of \u003cem\u003eH. pylori\u003c/em\u003e infections among Sudanese patients diagnosed with colon polyps and colon cancer and to correlate between its presence and the type of the lesions.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003eSample and data collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis is a preliminary, descriptive study aimed to investigate the frequency of \u003cem\u003eH. pylori\u003c/em\u003e infections among Sudanese patients diagnosed with colon cancer. Data were collected from 69 patients attended the National Laboratory and Alrahma Laboratory between February-June 2017.\u003c/p\u003e\n\u003cp\u003eDuring colonoscopy, presence of abnormal tissue, such as clumps of cells; polyps, formed on the inside of the colon, the pathologists obtained colon biopsies. Colon biopsies were fixed with formalin and processed into paraffin embedded blocks. Formalin-fixed paraffin blocks were used for the immunohistochemical detection of \u003cem\u003eH. pylori\u003c/em\u003e. Ethical approval was previously obtained by the pathologists of each hospital before colon polyps\u0026rsquo; biopsies were taken.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of the formalin fixed paraffin blocks\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFour sections from each formalin-fixed paraffin block to increase detection sensitivity were obtained with a thickness of 4\u0026micro;m using Rotary microtome (LEICA RM2125RT). All sections were de-waxed with two changes of Xylene for 3 minutes and then dehydrated in descending concentrations of Methanol starting from absolute Methanol through 90% and lastly, a concentration of 70% for 2 minutes in each concentration, and then washed using distilled water.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemistry diagnosis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eImmunohistochemistry diagnosis was performed on all the obtained sections. Known gastric sections containing \u003cem\u003eH. pylori\u003c/em\u003e infection was used as positive and negative controls; for the negative control the primary antibody incubation step was omitted. All sections were pretreated to retrieve antigens at 97\u003csup\u003eo\u003c/sup\u003eC for 10 minutes in citrate buffer solution, and then sections were blocked by 3% Hydrogen peroxide and absolute Methanol for 20 minutes at humidified chamber. Afterward, sections were blocked into Bovine serum Albumin (Thermo Fisher Scientific, Germany). A rabbit polyclonal antibody ULC3R (BioGenex, USA) (prepared from tissue culture supernatant diluted in PBS, pH 7.6 containing 5% BSA and 0.09% Sodium Azaide) against \u003cem\u003eH. pylori\u003c/em\u003e was applied for 40 minutes, then washed in buffer solution for 5 minutes. Then, a polymer solution was applied for 15 minutes, and washed in buffer solution for 5 minutes. Chromogen solution was added for 10 minutes, and washed in distilled water. Finally, Mayer\u0026rsquo;s Hematoxylin was added for 2 minutes, and then, sections were blued using running distilled water for 5 minutes. After bluing, sections were dehydrated, cleared, and mounted in DPX. Prepared sections were investigated microscopically by two experts\u0026rsquo; pathologists blindly without knowing the duplication of slides sections of each patient using X40 lens. Results were recorded into categories of positive and negative results; the dot like shape denoted the coccoid form of the organism as describe previously [36-39].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDescriptive data were analyzed using the Statistical Package for Social Science (SPSS-v20). Pearson Chi-square test was used to test the association of \u003cem\u003eH. pylori\u003c/em\u003e infection with the different types of lesions. A P value \u0026lt; 0.05 was considered as a statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eOf the 69 patients, there were 30 (43.5%) females and 39 (56.5%) males, their ages ranged from 21 to 80 years with a mean age 47.1\u0026plusmn;19.8. 44 (63.8%) patients were diagnosed with adenocarcinoma, 10 (14.5%) patients were colitis, and 15 (21.7%) patients were juvenile polyposis syndrome. No statistically significant was observed for the association of gender and the pathological condition of each patient, P value = 0.649. Out of the 69 patients, 16 (23.2%) patients were positive for \u003cem\u003eH. pylori\u003c/em\u003e infection. These 16 positive patients included; 13 (81.3%) patients diagnosed with adenocarcinoma and 3 (18.7%) patients diagnosed with juvenile polyps. The correlation between presence of \u003cem\u003eH. pylori \u003c/em\u003einfection and the histopathological condition of patients were positively correlated (P value 0.028) (Table 1).\u003c/p\u003e\n\u003cp\u003eIn respect to Immunohistochemistry diagnosis, the bacteria were prominent and easier to detect in the immune-stained sections in several patterns including organisms attached to the epithelial cells or within the superficial mucus. And in some cases, the bacteria were masked by inspissated mucus or being positioned flat and closely opposed to the epithelial surface. Regarding the morphological appearance of the organism; \u003cem\u003eH. pylori\u003c/em\u003e stained brown in color and take a dot and small curved shape in different sizes (Figure 1).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe exact role of\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;in the induction of colon cancer is still a debate between the scientific researcher communities; this is attributed to the controversial results obtained. In previous studies,\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;were linked to the development of gastric cancer [40], while others reported paradoxical results showing no association between \u003cem\u003eH. pylori \u003c/em\u003eand gastric cancer susceptibility [41-43]. However, reports from Sudan regarding the possible link between\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;and colon cancer are scarce. Therefore, in the present study, we examined the presence of\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;using immunohistochemistry technique on colon polyps and colon cancer lesions of Sudanese patients underwent colonoscopy.\u003c/p\u003e\n\u003cp\u003eThe results obtained from this study showed a positive correlation between the presence of\u0026nbsp;\u003cem\u003eH. pylori\u0026nbsp;\u003c/em\u003einfection and the histopathological diagnosis, as\u0026nbsp;\u003cem\u003eH. pylori\u0026nbsp;\u003c/em\u003ewas prevalent in higher frequency in patients diagnosed with adenocarcinoma compared to those diagnosed as juvenile polyposis syndrome, and the result was statistically significant. This result also agrees with studies conducted by Jones et al, and Grahn et al., they investigated the presence of\u0026nbsp;\u003cem\u003eH. pylori\u0026nbsp;\u003c/em\u003eamong 59 patients diagnosed with colon adenocarcinomas using immunohistochemistry technique, and 77 colon and rectum cancer patients using molecular technique, correspondingly [44, 45]. Jones et al., reported that\u0026nbsp;\u003cem\u003eH. pylori\u0026nbsp;\u003c/em\u003ewere detected in 10/59 adenocarcinoma cases which represent about 16.9% of the total cancer cases studied [44]. While, Grahn et al., showed that \u003cem\u003eH. pylori \u003c/em\u003ewere present in 27% of the patients; among the studied colon cancer, \u003cem\u003eH. pylori \u003c/em\u003ewere present in 11/42 (26%) patients [45].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAlthough, several studies failed to demonstrate any association between\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;and colon cancer, or even if this microorganism can colonize the colon [46-50]. This could only be attributed to the ability of demonstrating the \u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;bacteria, which was achieved by the aid of the immunohistochemistry technique that allowed a better localization of\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;within the various colon lesions of colitis, polyps, and adenocarcinoma included in this study.\u003c/p\u003e\n\u003cp\u003eInterestingly, several theories were proposed regarding the exact role by which\u0026nbsp;\u003cem\u003eH. pylori\u003c/em\u003e\u0026nbsp;induced colon cancer, one hypothesis is that colon cancer can be induced by toxins produced by \u003cem\u003eH. pylori\u003c/em\u003e; however, this theory was based only on serological data [43, 51-53]. Furthermore, some studies showed that colitis and colon cancer were also developed in experimental mice models infected with \u003cem\u003eH. hepaticus\u0026nbsp;\u003c/em\u003e[54]. Accordingly, the development of colon cancer seems most likely due to the interaction between toxins produced by the bacteria and the immune cells of the mice [54]. Therefore, the results we obtained from our study showing that \u003cem\u003eH. pylori \u003c/em\u003ewere present, nevertheless, it means that \u003cem\u003eH. pylori \u003c/em\u003einfection is responsible for the induction and the development of colon cancer, since the presence of \u003cem\u003eH. pylori \u003c/em\u003ecould be encountered as post-cancer incidence. This however, still requires more complicated experimental studies to investigate this hypothesis; yet, the role of \u003cem\u003eH. pylori \u003c/em\u003ecannot be excluded due to this hypothesis. Therefore, this preliminary report needs further advanced experimental investigations to enable the determination on the exact mechanisms by which \u003cem\u003eH. pylori \u003c/em\u003ecan induce colon cancer.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study was able to demonstrate the presence of \u003cem\u003eH. pylori \u003c/em\u003ein colon polyps and colon cancer using immunohistochemistry marker, besides the significant association in the presence of \u003cem\u003eH. pylori \u003c/em\u003ewith colon adenocarcinoma, indeed further studies are required to elaborate more in-depth about the exact role of \u003cem\u003eH. pylori \u003c/em\u003ein the development of colon cancer.\u003c/p\u003e"},{"header":"Limitations","content":"\u003cp\u003eIn this study the sample size studied was relatively small, but still a significant association was observed. A bigger sample size of colon cancer lesions and benign colon lesions; non cancer lesions, should be included in future studies to determine the significant association of \u003cem\u003eH. pylori\u003c/em\u003e with adenocarcinoma among the Sudanese patients diagnosed with colon cancer.\u0026nbsp;\u0026nbsp;\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eDNA: Deoxyribonucleic acid; CRC: Colorectal cancer; IM: Intestinal Metaplasia.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe study ethical clearance was obtained from University of Khartoum, Faculty of Medical Laboratory Sciences ethical review board. Informed consent was obtained from each participant prior to enrollment using writing and verbal informed consent.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eConsent to publish\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eNot Applicable.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe authors declare that they have no competing interest\u003cstrong\u003e\u003cem\u003es.\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eNot Applicable.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eAKM, NME, ZAA, FSA, EES and AMME provided conceptual framework for the project. AKM, NME, ZAA, FSA, ETA, NAM, RH, HAO and ESA participated in the preparation of the samples and performed the diagnosis. NSM, AF, EES, MSM, AA, and AMME performed the data analysis and guidance for data interpretation. AA, MSM, ETA, NAM, RH, ESA, AF, and NSM drafted the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe authors are grateful to the patients and the staff at the National Laboratory and Al-Rahma Laboratory for their continued collaboration and generous hospitality.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eAuthor\u0026rsquo;s Information\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003e\u003cstrong\u003eAziza K. Mohamed\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNazik M. Elhassan\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZahra A. Awhag\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFadwa S. Ali\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEman T. Ali\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan.\u003c/p\u003e\n\u003cp\u003e2 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, National University, Sudan, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNajwa A. Mhmoud\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e3 Mycetoma research center, Khartoum, Sudan.\u003c/p\u003e\n\u003cp\u003e4 Department of Microbiology and immunology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan. Email: \u003ca href=\"mailto:[email protected]\"\[email protected]\u003c/a\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEmmanuel E. Siddig\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e3 Mycetoma research center, University of Khartoum, Khartoum, Sudan.\u003c/p\u003e\n\u003cp\u003e5 Nile University, School of medicine, Khartoum, Sudan.\u003c/p\u003e\n\u003cp\u003e6 Department of Histopathology and Cytology, Alfarrabi College for Science and Technology, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRowa Hassan \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e3 Mycetoma Research Center, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEiman S. Ahmed \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e3 Mycetoma Research Center, University of Khartoum, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAzam Fattahi\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e7 Center for Research and Training in Skin Disease and Leprosy, Tehran University of Medical Sciences, Tehran, Iran. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAyman Ahmed\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e8 Department of Parasitology and Medical Entomology, Institute of Endemic Diseases, University of Khartoum, Sudan. Email: [email protected]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMohamed S. Muneer\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e9 Department of Neurology, Mayo Clinic, Jacksonville, FL, USA.\u003c/p\u003e\n\u003cp\u003e10 Department of Radiology, Mayo Clinic, Jacksonville, FL, USA\u003c/p\u003e\n\u003cp\u003e11 Department of Internal Medicine, Faculty of Medicine, University of Khartoum, Khartoum, Sudan. Email: [email protected]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHussam A. Osman\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e12 Biomedical Research Laboratory, Ahfad University for Women, Omdurman, Sudan.\u003c/p\u003e\n\u003cp\u003eEmail: [email protected] \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNouh S. Mohamed\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e12 Molecular Biology Department, Biotechnology Lab, Faculty of Pharmacy. Ahfad University for women, Khartoum Sudan.\u003c/p\u003e\n\u003cp\u003e13 Department of Parasitology and Medical Entomology, Faculty of Medicine, Sinnar University, Sinnar, Sudan.\u003c/p\u003e\n\u003cp\u003e14 Department of Parasitology and Medical Entomology, Faculty of Medical Laboratory Sciences, Nile University, Khartoum, Sudan. Email: [email protected].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAli MM. Edris\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan.\u003c/p\u003e\n\u003cp\u003e15 Faculty of Applied Medical Sciences, University of Bisha, Bisha, Kingdom of Saudi Arabia. Email: [email protected].\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eJemal A, Siegel R, Xu J, Ward E: \u003cstrong\u003eCancer statistics, 2010.\u003c/strong\u003e \u003cem\u003eCA: a cancer journal for clinicians \u003c/em\u003e2010, \u003cstrong\u003e60:\u003c/strong\u003e277-300.\u003c/li\u003e\n\u003cli\u003eHawkins NJ, Ward RL: \u003cstrong\u003eSporadic colorectal cancers with microsatellite instability and their possible origin in hyperplastic polyps and serrated adenomas.\u003c/strong\u003e \u003cem\u003eJournal of the National Cancer Institute \u003c/em\u003e2001, \u003cstrong\u003e93:\u003c/strong\u003e1307-1313.\u003c/li\u003e\n\u003cli\u003eJass JR: \u003cstrong\u003eHyperplastic-like polyps as precursors of microsatellite-unstable colorectal cancer.\u003c/strong\u003e \u003cem\u003eAmerican journal of clinical pathology \u003c/em\u003e2003, \u003cstrong\u003e119:\u003c/strong\u003e773-775.\u003c/li\u003e\n\u003cli\u003eKahi CJ, 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Passaro D, Figer A, Niv Y, Pitlik S, Samra Z, Koren R, Yahav J: \u003cstrong\u003eRelationship between Helicobacter pylori CagA status and colorectal cancer.\u003c/strong\u003e \u003cem\u003eThe American journal of gastroenterology \u003c/em\u003e2001, \u003cstrong\u003e96:\u003c/strong\u003e3406.\u003c/li\u003e\n\u003cli\u003eFireman Z, Trost L, Kopelman Y, Segal A, Sternberg A: \u003cstrong\u003eHelicobacter pylori: seroprevalence and colorectal cancer.\u003c/strong\u003e \u003cem\u003eThe Israel Medical Association journal: IMAJ \u003c/em\u003e2000, \u003cstrong\u003e2:\u003c/strong\u003e6-9.\u003c/li\u003e\n\u003cli\u003eMizuno S, Morita Y, Inui T, Asakawa A, Ueno N, Ando T, Kato H, Uchida M, Yoshikawa T, Inui AJIjoc: \u003cstrong\u003eHelicobacter pylori infection is associated with colon adenomatous polyps detected by high‐resolution colonoscopy.\u003c/strong\u003e 2005, \u003cstrong\u003e117:\u003c/strong\u003e1058-1059.\u003c/li\u003e\n\u003cli\u003eJones M, Helliwell P, Pritchard C, Tharakan J, Mathew JJWjoso: \u003cstrong\u003eHelicobacter pylori in colorectal neoplasms: is there an aetiological relationship?\u003c/strong\u003e 2007, \u003cstrong\u003e5:\u003c/strong\u003e51.\u003c/li\u003e\n\u003cli\u003eGrahn N, Hmani-Aifa M, Frans\u0026eacute;n K, S\u0026ouml;derkvist P, Monstein H-JJJomm: \u003cstrong\u003eMolecular identification of Helicobacter DNA present in human colorectal adenocarcinomas by 16S rDNA PCR amplification and pyrosequencing analysis.\u003c/strong\u003e 2005, \u003cstrong\u003e54:\u003c/strong\u003e1031-1035.\u003c/li\u003e\n\u003cli\u003eLimburg PJ, Stolzenberg-Solomon RZ, Colbert LH, Perez-Perez GI, Blaser MJ, Taylor PR, Virtamo J, Albanes D: \u003cstrong\u003eHelicobacter pylori seropositivity and colorectal cancer risk: a prospective study of male smokers.\u003c/strong\u003e \u003cem\u003eCancer Epidemiology Prevention Biomarkers \u003c/em\u003e2002, \u003cstrong\u003e11:\u003c/strong\u003e1095-1099.\u003c/li\u003e\n\u003cli\u003eMoss SF, Neugut AI, Garbowski GC, Wang S, Treat MR, Forde KA: \u003cstrong\u003eHelicobacter pylori seroprevalence and colorectal neoplasia: evidence against an association.\u003c/strong\u003e \u003cem\u003eJNCI: Journal of the National Cancer Institute \u003c/em\u003e1995, \u003cstrong\u003e87:\u003c/strong\u003e762-763.\u003c/li\u003e\n\u003cli\u003eSiddheshwar R, Muhammad K, Gray J, Kelly S: \u003cstrong\u003eSeroprevalence of Helicobacter pylori in patients with colorectal polyps and colorectal carcinoma.\u003c/strong\u003e \u003cem\u003eThe American journal of gastroenterology \u003c/em\u003e2001, \u003cstrong\u003e96:\u003c/strong\u003e84.\u003c/li\u003e\n\u003cli\u003eLuzza F, Maletta M, Imeneo M, Monteleone G, Marasco R, Biancone L, Pallone F: \u003cstrong\u003eEvidence against colonic mucosa colonisation by Helicobacter pylori. Lack of a specific antibody response in homogenates of rectal endoscopic biopsies.\u003c/strong\u003e \u003cem\u003eThe Italian journal of gastroenterology \u003c/em\u003e1996, \u003cstrong\u003e28:\u003c/strong\u003e447-451.\u003c/li\u003e\n\u003cli\u003eBell S, Chisholm S, Owen R, Borriello S, Kamm M: \u003cstrong\u003eEvaluation of Helicobacter species in inflammatory bowel disease.\u003c/strong\u003e \u003cem\u003eAlimentary pharmacology therapeutics \u003c/em\u003e2003, \u003cstrong\u003e18:\u003c/strong\u003e481-486.\u003c/li\u003e\n\u003cli\u003eShmuely H, Passaro D, Figer A, Niv Y, Pitlik S, Samra Z, Koren R, Yahav JJTAjog: \u003cstrong\u003eRelationship between Helicobacter pylori CagA status and colorectal cancer.\u003c/strong\u003e 2001, \u003cstrong\u003e96:\u003c/strong\u003e3406.\u003c/li\u003e\n\u003cli\u003eFireman Z, Trost L, Kopelman Y, Segal A, Sternberg AJTIMAjI: \u003cstrong\u003eHelicobacter pylori: seroprevalence and colorectal cancer.\u003c/strong\u003e 2000, \u003cstrong\u003e2:\u003c/strong\u003e6-9.\u003c/li\u003e\n\u003cli\u003eHsu W-Y, Lin C-H, Lin C-C, Sung F-C, Hsu C-P, Kao C-H: \u003cstrong\u003eThe relationship between Helicobacter pylori and cancer risk.\u003c/strong\u003e \u003cem\u003eEuropean journal of internal medicine \u003c/em\u003e2014, \u003cstrong\u003e25:\u003c/strong\u003e235-240.\u003c/li\u003e\n\u003cli\u003eBell S, Chisholm S, Owen R, Borriello S, Kamm MJAp, therapeutics: \u003cstrong\u003eEvaluation of Helicobacter species in inflammatory bowel disease.\u003c/strong\u003e 2003, \u003cstrong\u003e18:\u003c/strong\u003e481-486.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003e\u003cstrong\u003eTable 1:\u003c/strong\u003e Shows the correlation between gender, immunohistochemistry detection of \u003cem\u003eH. pylori\u003c/em\u003e with the histopathological diagnosis.\u003c/p\u003e\n\u003ctable border=\"1\" width=\"636\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"407\"\u003e\n\u003cp\u003e\u003cstrong\u003eHistopathological Diagnosis\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"84\"\u003e\n\u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"61\"\u003e\n\u003cp\u003e\u003cstrong\u003eP value\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e\u003cstrong\u003eAdenocarcinoma\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e\u003cstrong\u003eJuvenile Polyposis Syndrome\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e\u003cstrong\u003eColitis\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" width=\"636\"\u003e\n\u003cp\u003e\u003cstrong\u003eGender\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eMale\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e23 (58.9%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e10 (25.6%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e6 (15.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e39 (56.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"3\" width=\"61\"\u003e\n\u003cp\u003e0.649\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eFemale\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e21 (70.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e5 (16.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e4 (13.3%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e30 (43.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eTotal\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp; 44 (63.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e15 (21.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e10 (14.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e69 (100%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" width=\"636\"\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemistry of \u003cem\u003eH. Pylori\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eNegative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e31 (58.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e12 (22.6%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e10 (18.9%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e53 (76.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"3\" width=\"61\"\u003e\n\u003cp\u003e0.028\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003ePositive\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e13 (81.3%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp; 3 (18.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e0 (0.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e16 (23.2%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eTotal\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"126\"\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp; 44 (63.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"198\"\u003e\n\u003cp\u003e15 (21.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"83\"\u003e\n\u003cp\u003e10 (14.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e69 (100%)\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"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Colon polyps, Colon cancer, Helicobacter pylori, Immunohistochemistry","lastPublishedDoi":"10.21203/rs.3.rs-31738/v3","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-31738/v3","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjectives: \u003c/strong\u003eInfection with the bacteria \u003cem\u003eHelicobacter pylori \u003c/em\u003ehas been classified as class one carcinogen associated with increasing susceptibility of gastritis and gastric carcinoma. This study is aiming at investigating the prevalence of \u003cem\u003eH. pylori\u003c/em\u003e among colon polyps and colon cancer patients. A descriptive cross-sectional hospital-based study was conducted between February and June 2017. Sixty-nine formalin-fixed paraffin blocks collected from colon polyps and colon cancer patients to detect \u003cem\u003eH. pylori\u003c/em\u003e using immunohistochemistry technique. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Of the Sixty-nine patients included in the study, 39 (56.5%) males and 30 (43.5%) were females, their age ranged from 21 to 80 years with a mean age of 47.1±19.7. Of the 69 colon polyps and colon cancer patients, 44 (63.8%) were diagnosed as adenocarcinoma, 10 (14.5%) colitis, 15 (21.7%) juvenile polyposis syndrome. The results of immunohistochemistry technique showed the presence of 16 (23.2%) positive patients for \u003cem\u003eH. pylori\u003c/em\u003e infection. Of these 16, 13 (81.3%) patients were diagnosed with adenocarcinoma and 3 (18.7%) patients were diagnosed with juvenile polyps. The results of \u003cem\u003eH. pylori\u003c/em\u003e detection among the different colon polyps and colon cancer patients were showing a statistically significant association for \u003cem\u003eH. pylori\u003c/em\u003e infection and adenocarcinoma, P value 0.028.\u003c/p\u003e","manuscriptTitle":"Prevalence of Helicobacter pylori among Sudanese patients diagnosed with colon polyps and colon cancer using immunohistochemistry technique.","msid":"","msnumber":"","nonDraftVersions":[{"code":3,"date":"2020-06-25 18:42:24","doi":"10.21203/rs.3.rs-31738/v3","editorialEvents":[{"type":"communityComments","content":0},{"type":"checksComplete","content":"","date":"2020-06-25T12:00:00+00:00","index":"","fulltext":""},{"type":"decision","content":"Accept","date":"2020-06-25T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}},{"code":2,"date":"2020-06-23 17:40:52","doi":"10.21203/rs.3.rs-31738/v2","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Minor revision","date":"2020-06-23T12:00:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2020-06-22T12:00:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2020-06-21T12:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2020-06-21T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2020-05-28 21:38:46","doi":"10.21203/rs.3.rs-31738/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2020-06-14T12:00:00+00:00","index":1,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2020-06-14T12:00:00+00:00","index":1,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"decision","content":"Major revision","date":"2020-06-14T12:00:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2020-05-26T12:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2020-05-26T12:00:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2020-05-26T12:00:00+00:00","index":"","fulltext":""},{"type":"reviewersInvited","content":"","date":"2020-05-26T12:00:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"","date":"2020-05-24T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e13e64e3-970e-4171-a675-55e503754f3a","owner":[],"postedDate":"June 25th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":131254,"name":"Cancer Biology"}],"tags":[],"updatedAt":"2020-07-12T15:34:17+00:00","versionOfRecord":{"articleIdentity":"rs-31738","link":"https://doi.org/10.1186/s13104-020-05159-2","journal":{"identity":"bmc-research-notes","isVorOnly":false,"title":"BMC Research Notes"},"publishedOn":"2020-07-06 12:00:00","publishedOnDateReadable":"July 6th, 2020"},"versionCreatedAt":"2020-06-25 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