Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study

Kadukkai maathirai (Indian herbal drug) prevents... | F1000Research "use strict";function _typeof(t){return(_typeof="function"==typeof Symbol&&"symbol"==typeof Symbol.iterator?function(t){return typeof t}:function(t){return t&&"function"==typeof Symbol&&t.constructor===Symbol&&t!==Symbol.prototype?"symbol":typeof t})(t)}!function(){var t=function(){var t,e,o=[],n=window,r=n;for(;r;){try{if(r.frames.__tcfapiLocator){t=r;break}}catch(t){}if(r===n.top)break;r=r.parent}t||(!function t(){var e=n.document,o=!!n.frames.__tcfapiLocator;if(!o)if(e.body){var r=e.createElement("iframe");r.style.cssText="display:none",r.name="__tcfapiLocator",e.body.appendChild(r)}else setTimeout(t,5);return!o}(),n.__tcfapi=function(){for(var t=arguments.length,n=new Array(t),r=0;r 3&&2===parseInt(n[1],10)&&"boolean"==typeof n[3]&&(e=n[3],"function"==typeof n[2]&&n[2]("set",!0)):"ping"===n[0]?"function"==typeof n[2]&&n[2]({gdprApplies:e,cmpLoaded:!1,cmpStatus:"stub"}):o.push(n)},n.addEventListener("message",(function(t){var e="string"==typeof t.data,o={};if(e)try{o=JSON.parse(t.data)}catch(t){}else o=t.data;var n="object"===_typeof(o)&&null!==o?o.__tcfapiCall:null;n&&window.__tcfapi(n.command,n.version,(function(o,r){var a={__tcfapiReturn:{returnValue:o,success:r,callId:n.callId}};t&&t.source&&t.source.postMessage&&t.source.postMessage(e?JSON.stringify(a):a,"*")}),n.parameter)}),!1))};"undefined"!=typeof module?module.exports=t:t()}(); dataLayer = dataLayer || []; // Standard GTM initialization - Google Consent Mode handles consent automatically (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+ '>m_auth=hzk0Vc3qFsQYhCrIoHz68A>m_preview=env-1>m_cookies_win=x';f.parentNode.insertBefore(j,f); })(window,document,'script','dataLayer','GTM-MWFK8L5J'); ;window.NREUM||(NREUM={});NREUM.init={distributed_tracing:{enabled:true},privacy:{cookies_enabled:true},ajax:{deny_list:["bam.nr-data.net"]}}; ;NREUM.loader_config={accountID:"438030",trustKey:"438030",agentID:"772317073",licenseKey:"97f8f67f26",applicationID:"772317073"} ;NREUM.info={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net",licenseKey:"97f8f67f26",applicationID:"772317073",sa:1} ;/*! For license information please see nr-loader-spa-1.236.0.min.js.LICENSE.txt */ (()=>{"use strict";var e,t,r={5763:(e,t,r)=>{r.d(t,{P_:()=>l,Mt:()=>g,C5:()=>s,DL:()=>v,OP:()=>T,lF:()=>D,Yu:()=>y,Dg:()=>h,CX:()=>c,GE:()=>b,sU:()=>_});var n=r(8632),i=r(9567);const o={beacon:n.ce.beacon,errorBeacon:n.ce.errorBeacon,licenseKey:void 0,applicationID:void 0,sa:void 0,queueTime:void 0,applicationTime:void 0,ttGuid:void 0,user:void 0,account:void 0,product:void 0,extra:void 0,jsAttributes:{},userAttributes:void 0,atts:void 0,transactionName:void 0,tNamePlain:void 0},a={};function s(e){if(!e)throw new Error("All info objects require an agent identifier!");if(!a[e])throw new Error("Info for ".concat(e," was never set"));return a[e]}function c(e,t){if(!e)throw new Error("All info objects require an agent identifier!");a[e]=(0,i.D)(t,o),(0,n.Qy)(e,a[e],"info")}var u=r(7056);const d=()=>{const e={blockSelector:"[data-nr-block]",maskInputOptions:{password:!0}};return{allow_bfcache:!0,privacy:{cookies_enabled:!0},ajax:{deny_list:void 0,enabled:!0,harvestTimeSeconds:10},distributed_tracing:{enabled:void 0,exclude_newrelic_header:void 0,cors_use_newrelic_header:void 0,cors_use_tracecontext_headers:void 0,allowed_origins:void 0},session:{domain:void 0,expiresMs:u.oD,inactiveMs:u.Hb},ssl:void 0,obfuscate:void 0,jserrors:{enabled:!0,harvestTimeSeconds:10},metrics:{enabled:!0},page_action:{enabled:!0,harvestTimeSeconds:30},page_view_event:{enabled:!0},page_view_timing:{enabled:!0,harvestTimeSeconds:30,long_task:!1},session_trace:{enabled:!0,harvestTimeSeconds:10},harvest:{tooManyRequestsDelay:60},session_replay:{enabled:!1,harvestTimeSeconds:60,sampleRate:.1,errorSampleRate:.1,maskTextSelector:"*",maskAllInputs:!0,get blockClass(){return"nr-block"},get ignoreClass(){return"nr-ignore"},get maskTextClass(){return"nr-mask"},get blockSelector(){return e.blockSelector},set blockSelector(t){e.blockSelector+=",".concat(t)},get maskInputOptions(){return e.maskInputOptions},set maskInputOptions(t){e.maskInputOptions={...t,password:!0}}},spa:{enabled:!0,harvestTimeSeconds:10}}},f={};function l(e){if(!e)throw new Error("All configuration objects require an agent identifier!");if(!f[e])throw new Error("Configuration for ".concat(e," was never set"));return f[e]}function h(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");f[e]=(0,i.D)(t,d()),(0,n.Qy)(e,f[e],"config")}function g(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");var r=l(e);if(r){for(var n=t.split("."),i=0;i {r.d(t,{D:()=>i});var n=r(50);function i(e,t){try{if(!e||"object"!=typeof e)return(0,n.Z)("Setting a Configurable requires an object as input");if(!t||"object"!=typeof t)return(0,n.Z)("Setting a Configurable requires a model to set its initial properties");const r=Object.create(Object.getPrototypeOf(t),Object.getOwnPropertyDescriptors(t)),o=0===Object.keys(r).length?e:r;for(let a in o)if(void 0!==e[a])try{"object"==typeof e[a]&&"object"==typeof t[a]?r[a]=i(e[a],t[a]):r[a]=e[a]}catch(e){(0,n.Z)("An error occurred while setting a property of a Configurable",e)}return r}catch(e){(0,n.Z)("An error occured while setting a Configurable",e)}}},6818:(e,t,r)=>{r.d(t,{Re:()=>i,gF:()=>o,q4:()=>n});const n="1.236.0",i="PROD",o="CDN"},385:(e,t,r)=>{r.d(t,{FN:()=>a,IF:()=>u,Nk:()=>f,Tt:()=>s,_A:()=>o,il:()=>n,pL:()=>c,v6:()=>i,w1:()=>d});const n="undefined"!=typeof window&&!!window.document,i="undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self.navigator instanceof WorkerNavigator||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis.navigator instanceof WorkerNavigator),o=n?window:"undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis),a=""+o?.location,s=/iPad|iPhone|iPod/.test(navigator.userAgent),c=s&&"undefined"==typeof SharedWorker,u=(()=>{const e=navigator.userAgent.match(/Firefox[/\s](\d+\.\d+)/);return Array.isArray(e)&&e.length>=2?+e[1]:0})(),d=Boolean(n&&window.document.documentMode),f=!!navigator.sendBeacon},1117:(e,t,r)=>{r.d(t,{w:()=>o});var n=r(50);const i={agentIdentifier:"",ee:void 0};class o{constructor(e){try{if("object"!=typeof e)return(0,n.Z)("shared context requires an object as input");this.sharedContext={},Object.assign(this.sharedContext,i),Object.entries(e).forEach((e=>{let[t,r]=e;Object.keys(i).includes(t)&&(this.sharedContext[t]=r)}))}catch(e){(0,n.Z)("An error occured while setting SharedContext",e)}}}},8e3:(e,t,r)=>{r.d(t,{L:()=>d,R:()=>c});var n=r(2177),i=r(1284),o=r(4322),a=r(3325);const s={};function c(e,t){const r={staged:!1,priority:a.p[t]||0};u(e),s[e].get(t)||s[e].set(t,r)}function u(e){e&&(s[e]||(s[e]=new Map))}function d(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:"",t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:"feature";if(u(e),!e||!s[e].get(t))return a(t);s[e].get(t).staged=!0;const r=[...s[e]];function a(t){const r=e?n.ee.get(e):n.ee,a=o.X.handlers;if(r.backlog&&a){var s=r.backlog[t],c=a[t];if(c){for(var u=0;s&&u {let[t,r]=e;return r.staged}))&&(r.sort(((e,t)=>e[1].priority-t[1].priority)),r.forEach((e=>{let[t]=e;a(t)})))}function f(e,t){var r=e[1];(0,i.D)(t[r],(function(t,r){var n=e[0];if(r[0]===n){var i=r[1],o=e[3],a=e[2];i.apply(o,a)}}))}},2177:(e,t,r)=>{r.d(t,{c:()=>f,ee:()=>u});var n=r(8632),i=r(2210),o=r(1284),a=r(5763),s="nr@context";let c=(0,n.fP)();var u;function d(){}function f(e){return(0,i.X)(e,s,l)}function l(){return new d}function h(){u.aborted=!0,u.backlog={}}c.ee?u=c.ee:(u=function e(t,r){var n={},c={},f={},g=!1;try{g=16===r.length&&(0,a.OP)(r).isolatedBacklog}catch(e){}var p={on:b,addEventListener:b,removeEventListener:y,emit:v,get:x,listeners:w,context:m,buffer:A,abort:h,aborted:!1,isBuffering:E,debugId:r,backlog:g?{}:t&&"object"==typeof t.backlog?t.backlog:{}};return p;function m(e){return e&&e instanceof d?e:e?(0,i.X)(e,s,l):l()}function v(e,r,n,i,o){if(!1!==o&&(o=!0),!u.aborted||i){t&&o&&t.emit(e,r,n);for(var a=m(n),s=w(e),d=s.length,f=0;fn,p:()=>i});var n=r(2177).ee.get("handle");function i(e,t,r,i,o){o?(o.buffer([e],i),o.emit(e,t,r)):(n.buffer([e],i),n.emit(e,t,r))}},4322:(e,t,r)=>{r.d(t,{X:()=>o});var n=r(5546);o.on=a;var i=o.handlers={};function o(e,t,r,o){a(o||n.E,i,e,t,r)}function a(e,t,r,i,o){o||(o="feature"),e||(e=n.E);var a=t[o]=t[o]||{};(a[r]=a[r]||[]).push([e,i])}},3239:(e,t,r)=>{r.d(t,{bP:()=>s,iz:()=>c,m$:()=>a});var n=r(385);let i=!1,o=!1;try{const e={get passive(){return i=!0,!1},get signal(){return o=!0,!1}};n._A.addEventListener("test",null,e),n._A.removeEventListener("test",null,e)}catch(e){}function a(e,t){return i||o?{capture:!!e,passive:i,signal:t}:!!e}function s(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;window.addEventListener(e,t,a(r,n))}function c(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;document.addEventListener(e,t,a(r,n))}},4402:(e,t,r)=>{r.d(t,{Ht:()=>u,M:()=>c,Rl:()=>a,ky:()=>s});var n=r(385);const i="xxxxxxxx-xxxx-4xxx-yxxx-xxxxxxxxxxxx";function o(e,t){return e?15&e[t]:16*Math.random()|0}function a(){const e=n._A?.crypto||n._A?.msCrypto;let t,r=0;return e&&e.getRandomValues&&(t=e.getRandomValues(new Uint8Array(31))),i.split("").map((e=>"x"===e?o(t,++r).toString(16):"y"===e?(3&o()|8).toString(16):e)).join("")}function s(e){const t=n._A?.crypto||n._A?.msCrypto;let r,i=0;t&&t.getRandomValues&&(r=t.getRandomValues(new Uint8Array(31)));const a=[];for(var s=0;s {r.d(t,{Bq:()=>n,Hb:()=>o,oD:()=>i});const n="NRBA",i=144e5,o=18e5},7894:(e,t,r)=>{function n(){return Math.round(performance.now())}r.d(t,{z:()=>n})},7243:(e,t,r)=>{r.d(t,{e:()=>o});var n=r(385),i={};function o(e){if(e in i)return i[e];if(0===(e||"").indexOf("data:"))return{protocol:"data"};let t;var r=n._A?.location,o={};if(n.il)t=document.createElement("a"),t.href=e;else try{t=new URL(e,r.href)}catch(e){return o}o.port=t.port;var a=t.href.split("://");!o.port&&a[1]&&(o.port=a[1].split("/")[0].split("@").pop().split(":")[1]),o.port&&"0"!==o.port||(o.port="https"===a[0]?"443":"80"),o.hostname=t.hostname||r.hostname,o.pathname=t.pathname,o.protocol=a[0],"/"!==o.pathname.charAt(0)&&(o.pathname="/"+o.pathname);var s=!t.protocol||":"===t.protocol||t.protocol===r.protocol,c=t.hostname===r.hostname&&t.port===r.port;return o.sameOrigin=s&&(!t.hostname||c),"/"===o.pathname&&(i[e]=o),o}},50:(e,t,r)=>{function n(e,t){"function"==typeof console.warn&&(console.warn("New Relic: ".concat(e)),t&&console.warn(t))}r.d(t,{Z:()=>n})},2587:(e,t,r)=>{r.d(t,{N:()=>c,T:()=>u});var n=r(2177),i=r(5546),o=r(8e3),a=r(3325);const s={stn:[a.D.sessionTrace],err:[a.D.jserrors,a.D.metrics],ins:[a.D.pageAction],spa:[a.D.spa],sr:[a.D.sessionReplay,a.D.sessionTrace]};function c(e,t){const r=n.ee.get(t);e&&"object"==typeof e&&(Object.entries(e).forEach((e=>{let[t,n]=e;void 0===u[t]&&(s[t]?s[t].forEach((e=>{n?(0,i.p)("feat-"+t,[],void 0,e,r):(0,i.p)("block-"+t,[],void 0,e,r),(0,i.p)("rumresp-"+t,[Boolean(n)],void 0,e,r)})):n&&(0,i.p)("feat-"+t,[],void 0,void 0,r),u[t]=Boolean(n))})),Object.keys(s).forEach((e=>{void 0===u[e]&&(s[e]?.forEach((t=>(0,i.p)("rumresp-"+e,[!1],void 0,t,r))),u[e]=!1)})),(0,o.L)(t,a.D.pageViewEvent))}const u={}},2210:(e,t,r)=>{r.d(t,{X:()=>i});var n=Object.prototype.hasOwnProperty;function i(e,t,r){if(n.call(e,t))return e[t];var i=r();if(Object.defineProperty&&Object.keys)try{return Object.defineProperty(e,t,{value:i,writable:!0,enumerable:!1}),i}catch(e){}return e[t]=i,i}},1284:(e,t,r)=>{r.d(t,{D:()=>n});const n=(e,t)=>Object.entries(e||{}).map((e=>{let[r,n]=e;return t(r,n)}))},4351:(e,t,r)=>{r.d(t,{P:()=>o});var n=r(2177);const i=()=>{const e=new WeakSet;return(t,r)=>{if("object"==typeof r&&null!==r){if(e.has(r))return;e.add(r)}return r}};function o(e){try{return JSON.stringify(e,i())}catch(e){try{n.ee.emit("internal-error",[e])}catch(e){}}}},3960:(e,t,r)=>{r.d(t,{K:()=>a,b:()=>o});var n=r(3239);function i(){return"undefined"==typeof document||"complete"===document.readyState}function o(e,t){if(i())return e();(0,n.bP)("load",e,t)}function a(e){if(i())return e();(0,n.iz)("DOMContentLoaded",e)}},8632:(e,t,r)=>{r.d(t,{EZ:()=>u,Qy:()=>c,ce:()=>o,fP:()=>a,gG:()=>d,mF:()=>s});var n=r(7894),i=r(385);const o={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net"};function a(){return i._A.NREUM||(i._A.NREUM={}),void 0===i._A.newrelic&&(i._A.newrelic=i._A.NREUM),i._A.NREUM}function s(){let e=a();return e.o||(e.o={ST:i._A.setTimeout,SI:i._A.setImmediate,CT:i._A.clearTimeout,XHR:i._A.XMLHttpRequest,REQ:i._A.Request,EV:i._A.Event,PR:i._A.Promise,MO:i._A.MutationObserver,FETCH:i._A.fetch}),e}function c(e,t,r){let i=a();const o=i.initializedAgents||{},s=o[e]||{};return Object.keys(s).length||(s.initializedAt={ms:(0,n.z)(),date:new Date}),i.initializedAgents={...o,[e]:{...s,[r]:t}},i}function u(e,t){a()[e]=t}function d(){return function(){let e=a();const t=e.info||{};e.info={beacon:o.beacon,errorBeacon:o.errorBeacon,...t}}(),function(){let e=a();const t=e.init||{};e.init={...t}}(),s(),function(){let e=a();const t=e.loader_config||{};e.loader_config={...t}}(),a()}},7956:(e,t,r)=>{r.d(t,{N:()=>i});var n=r(3239);function i(e){let t=arguments.length>1&&void 0!==arguments[1]&&arguments[1],r=arguments.length>2?arguments[2]:void 0,i=arguments.length>3?arguments[3]:void 0;return void(0,n.iz)("visibilitychange",(function(){if(t)return void("hidden"==document.visibilityState&&e());e(document.visibilityState)}),r,i)}},1214:(e,t,r)=>{r.d(t,{em:()=>v,u5:()=>N,QU:()=>S,_L:()=>I,Gm:()=>L,Lg:()=>M,gy:()=>U,BV:()=>Q,Kf:()=>ee});var n=r(2177);const i="nr@original";var o=Object.prototype.hasOwnProperty,a=!1;function s(e,t){return e||(e=n.ee),r.inPlace=function(e,t,n,i,o){n||(n="");var a,s,c,u="-"===n.charAt(0);for(c=0;c 2?n-2:0),o=2;o {r(A[T],e,w),r(E[T],e,w)})),r(l._A,"fetch",y),t.on(y+"end",(function(e,r){var n=this;if(r){var i=r.headers.get("content-length");null!==i&&(n.rxSize=i),t.emit(y+"done",[null,r],n)}else t.emit(y+"done",[e],n)})),t}const O={},j=["pushState","replaceState"];function S(e){const t=function(e){return(e||n.ee).get("history")}(e);return!l.il||O[t.debugId]++||(O[t.debugId]=1,s(t).inPlace(window.history,j,"-")),t}var P=r(3239);const C={},R=["appendChild","insertBefore","replaceChild"];function I(e){const t=function(e){return(e||n.ee).get("jsonp")}(e);if(!l.il||C[t.debugId])return t;C[t.debugId]=!0;var r=s(t),i=/[?&](?:callback|cb)=([^&#]+)/,o=/(.*)\.([^.]+)/,a=/^(\w+)(\.|$)(.*)$/;function c(e,t){var r=e.match(a),n=r[1],i=r[3];return i?c(i,t[n]):t[n]}return r.inPlace(Node.prototype,R,"dom-"),t.on("dom-start",(function(e){!function(e){if(!e||"string"!=typeof e.nodeName||"script"!==e.nodeName.toLowerCase())return;if("function"!=typeof e.addEventListener)return;var n=(a=e.src,s=a.match(i),s?s[1]:null);var a,s;if(!n)return;var u=function(e){var t=e.match(o);if(t&&t.length>=3)return{key:t[2],parent:c(t[1],window)};return{key:e,parent:window}}(n);if("function"!=typeof u.parent[u.key])return;var d={};function f(){t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}function l(){t.emit("jsonp-error",[],d),t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}r.inPlace(u.parent,[u.key],"cb-",d),e.addEventListener("load",f,(0,P.m$)(!1)),e.addEventListener("error",l,(0,P.m$)(!1)),t.emit("new-jsonp",[e.src],d)}(e[0])})),t}var k=r(5763);const H={};function L(e){const t=function(e){return(e||n.ee).get("mutation")}(e);if(!l.il||H[t.debugId])return t;H[t.debugId]=!0;var r=s(t),i=k.Yu.MO;return i&&(window.MutationObserver=function(e){return this instanceof i?new i(r(e,"fn-")):i.apply(this,arguments)},MutationObserver.prototype=i.prototype),t}const z={};function M(e){const t=function(e){return(e||n.ee).get("promise")}(e);if(z[t.debugId])return t;z[t.debugId]=!0;var r=n.c,o=s(t),a=k.Yu.PR;return a&&function(){function e(r){var n=t.context(),i=o(r,"executor-",n,null,!1);const s=Reflect.construct(a,[i],e);return t.context(s).getCtx=function(){return n},s}l._A.Promise=e,Object.defineProperty(e,"name",{value:"Promise"}),e.toString=function(){return a.toString()},Object.setPrototypeOf(e,a),["all","race"].forEach((function(r){const n=a[r];e[r]=function(e){let i=!1;[...e||[]].forEach((e=>{this.resolve(e).then(a("all"===r),a(!1))}));const o=n.apply(this,arguments);return o;function a(e){return function(){t.emit("propagate",[null,!i],o,!1,!1),i=i||!e}}}})),["resolve","reject"].forEach((function(r){const n=a[r];e[r]=function(e){const r=n.apply(this,arguments);return e!==r&&t.emit("propagate",[e,!0],r,!1,!1),r}})),e.prototype=a.prototype;const n=a.prototype.then;a.prototype.then=function(){var e=this,i=r(e);i.promise=e;for(var a=arguments.length,s=new Array(a),c=0;c e())),t};function m(e,t){i.inPlace(t,["onreadystatechange"],"fn-",E)}function b(){var e=this,t=r.context(e);e.readyState>3&&!t.resolved&&(t.resolved=!0,r.emit("xhr-resolved",[],e)),i.inPlace(e,f,"fn-",E)}if(function(e,t){for(var r in e)t[r]=e[r]}(o,p),p.prototype=o.prototype,i.inPlace(p.prototype,J,"-xhr-",E),r.on("send-xhr-start",(function(e,t){m(e,t),function(e){h.push(e),a&&(y?y.then(A):u?u(A):(w=-w,x.data=w))}(t)})),r.on("open-xhr-start",m),a){var y=c&&c.resolve();if(!u&&!c){var w=1,x=document.createTextNode(w);new a(A).observe(x,{characterData:!0})}}else t.on("fn-end",(function(e){e[0]&&e[0].type===d||A()}));function A(){for(var e=0;e {r.d(t,{t:()=>n});const n=r(3325).D.ajax},6660:(e,t,r)=>{r.d(t,{A:()=>i,t:()=>n});const n=r(3325).D.jserrors,i="nr@seenError"},3081:(e,t,r)=>{r.d(t,{gF:()=>o,mY:()=>i,t9:()=>n,vz:()=>s,xS:()=>a});const n=r(3325).D.metrics,i="sm",o="cm",a="storeSupportabilityMetrics",s="storeEventMetrics"},4649:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageAction},7633:(e,t,r)=>{r.d(t,{Dz:()=>i,OJ:()=>a,qw:()=>o,t9:()=>n});const n=r(3325).D.pageViewEvent,i="firstbyte",o="domcontent",a="windowload"},9251:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageViewTiming},3614:(e,t,r)=>{r.d(t,{BST_RESOURCE:()=>i,END:()=>s,FEATURE_NAME:()=>n,FN_END:()=>u,FN_START:()=>c,PUSH_STATE:()=>d,RESOURCE:()=>o,START:()=>a});const n=r(3325).D.sessionTrace,i="bstResource",o="resource",a="-start",s="-end",c="fn"+a,u="fn"+s,d="pushState"},7836:(e,t,r)=>{r.d(t,{BODY:()=>A,CB_END:()=>E,CB_START:()=>u,END:()=>x,FEATURE_NAME:()=>i,FETCH:()=>_,FETCH_BODY:()=>v,FETCH_DONE:()=>m,FETCH_START:()=>p,FN_END:()=>c,FN_START:()=>s,INTERACTION:()=>l,INTERACTION_API:()=>d,INTERACTION_EVENTS:()=>o,JSONP_END:()=>b,JSONP_NODE:()=>g,JS_TIME:()=>T,MAX_TIMER_BUDGET:()=>a,REMAINING:()=>f,SPA_NODE:()=>h,START:()=>w,originalSetTimeout:()=>y});var n=r(5763);const i=r(3325).D.spa,o=["click","submit","keypress","keydown","keyup","change"],a=999,s="fn-start",c="fn-end",u="cb-start",d="api-ixn-",f="remaining",l="interaction",h="spaNode",g="jsonpNode",p="fetch-start",m="fetch-done",v="fetch-body-",b="jsonp-end",y=n.Yu.ST,w="-start",x="-end",A="-body",E="cb"+x,T="jsTime",_="fetch"},5938:(e,t,r)=>{r.d(t,{W:()=>o});var n=r(5763),i=r(2177);class o{constructor(e,t,r){this.agentIdentifier=e,this.aggregator=t,this.ee=i.ee.get(e,(0,n.OP)(this.agentIdentifier).isolatedBacklog),this.featureName=r,this.blocked=!1}}},9144:(e,t,r)=>{r.d(t,{j:()=>m});var n=r(3325),i=r(5763),o=r(5546),a=r(2177),s=r(7894),c=r(8e3),u=r(3960),d=r(385),f=r(50),l=r(3081),h=r(8632);function g(){const e=(0,h.gG)();["setErrorHandler","finished","addToTrace","inlineHit","addRelease","addPageAction","setCurrentRouteName","setPageViewName","setCustomAttribute","interaction","noticeError","setUserId"].forEach((t=>{e[t]=function(){for(var r=arguments.length,n=new Array(r),i=0;i 1?r-1:0),i=1;i {e.exposed&&e.api[t]&&o.push(e.api[t](...n))})),o.length>1?o:o[0]}(t,...n)}}))}var p=r(2587);function m(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:{},m=arguments.length>2?arguments[2]:void 0,v=arguments.length>3?arguments[3]:void 0,{init:b,info:y,loader_config:w,runtime:x={loaderType:m},exposed:A=!0}=t;const E=(0,h.gG)();y||(b=E.init,y=E.info,w=E.loader_config),(0,i.Dg)(e,b||{}),(0,i.GE)(e,w||{}),(0,i.sU)(e,x),y.jsAttributes??={},d.v6&&(y.jsAttributes.isWorker=!0),(0,i.CX)(e,y),g();const T=function(e,t){t||(0,c.R)(e,"api");const h={};var g=a.ee.get(e),p=g.get("tracer"),m="api-",v=m+"ixn-";function b(t,r,n,o){const a=(0,i.C5)(e);return null===r?delete a.jsAttributes[t]:(0,i.CX)(e,{...a,jsAttributes:{...a.jsAttributes,[t]:r}}),x(m,n,!0,o||null===r?"session":void 0)(t,r)}function y(){}["setErrorHandler","finished","addToTrace","inlineHit","addRelease"].forEach((e=>h[e]=x(m,e,!0,"api"))),h.addPageAction=x(m,"addPageAction",!0,n.D.pageAction),h.setCurrentRouteName=x(m,"routeName",!0,n.D.spa),h.setPageViewName=function(t,r){if("string"==typeof t)return"/"!==t.charAt(0)&&(t="/"+t),(0,i.OP)(e).customTransaction=(r||"http://custom.transaction")+t,x(m,"setPageViewName",!0)()},h.setCustomAttribute=function(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2];if("string"==typeof e){if(["string","number"].includes(typeof t)||null===t)return b(e,t,"setCustomAttribute",r);(0,f.Z)("Failed to execute setCustomAttribute.\nNon-null value must be a string or number type, but a type of was provided."))}else(0,f.Z)("Failed to execute setCustomAttribute.\nName must be a string type, but a type of was provided."))},h.setUserId=function(e){if("string"==typeof e||null===e)return b("enduser.id",e,"setUserId",!0);(0,f.Z)("Failed to execute setUserId.\nNon-null value must be a string type, but a type of was provided."))},h.interaction=function(){return(new y).get()};var w=y.prototype={createTracer:function(e,t){var r={},i=this,a="function"==typeof t;return(0,o.p)(v+"tracer",[(0,s.z)(),e,r],i,n.D.spa,g),function(){if(p.emit((a?"":"no-")+"fn-start",[(0,s.z)(),i,a],r),a)try{return t.apply(this,arguments)}catch(e){throw p.emit("fn-err",[arguments,this,"string"==typeof e?new Error(e):e],r),e}finally{p.emit("fn-end",[(0,s.z)()],r)}}}};function x(e,t,r,i){return function(){return(0,o.p)(l.xS,["API/"+t+"/called"],void 0,n.D.metrics,g),i&&(0,o.p)(e+t,[(0,s.z)(),...arguments],r?null:this,i,g),r?void 0:this}}function A(){r.e(439).then(r.bind(r,7438)).then((t=>{let{setAPI:r}=t;r(e),(0,c.L)(e,"api")})).catch((()=>(0,f.Z)("Downloading runtime APIs failed...")))}return["actionText","setName","setAttribute","save","ignore","onEnd","getContext","end","get"].forEach((e=>{w[e]=x(v,e,void 0,n.D.spa)})),h.noticeError=function(e,t){"string"==typeof e&&(e=new Error(e)),(0,o.p)(l.xS,["API/noticeError/called"],void 0,n.D.metrics,g),(0,o.p)("err",[e,(0,s.z)(),!1,t],void 0,n.D.jserrors,g)},d.il?(0,u.b)((()=>A()),!0):A(),h}(e,v);return(0,h.Qy)(e,T,"api"),(0,h.Qy)(e,A,"exposed"),(0,h.EZ)("activatedFeatures",p.T),T}},3325:(e,t,r)=>{r.d(t,{D:()=>n,p:()=>i});const n={ajax:"ajax",jserrors:"jserrors",metrics:"metrics",pageAction:"page_action",pageViewEvent:"page_view_event",pageViewTiming:"page_view_timing",sessionReplay:"session_replay",sessionTrace:"session_trace",spa:"spa"},i={[n.pageViewEvent]:1,[n.pageViewTiming]:2,[n.metrics]:3,[n.jserrors]:4,[n.ajax]:5,[n.sessionTrace]:6,[n.pageAction]:7,[n.spa]:8,[n.sessionReplay]:9}}},n={};function i(e){var t=n[e];if(void 0!==t)return t.exports;var o=n[e]={exports:{}};return r[e](o,o.exports,i),o.exports}i.m=r,i.d=(e,t)=>{for(var r in t)i.o(t,r)&&!i.o(e,r)&&Object.defineProperty(e,r,{enumerable:!0,get:t[r]})},i.f={},i.e=e=>Promise.all(Object.keys(i.f).reduce(((t,r)=>(i.f[r](e,t),t)),[])),i.u=e=>(({78:"page_action-aggregate",147:"metrics-aggregate",242:"session-manager",317:"jserrors-aggregate",348:"page_view_timing-aggregate",412:"lazy-feature-loader",439:"async-api",538:"recorder",590:"session_replay-aggregate",675:"compressor",733:"session_trace-aggregate",786:"page_view_event-aggregate",873:"spa-aggregate",898:"ajax-aggregate"}[e]||e)+"."+{78:"ac76d497",147:"3dc53903",148:"1a20d5fe",242:"2a64278a",317:"49e41428",348:"bd6de33a",412:"2f55ce66",439:"30bd804e",538:"1b18459f",590:"cf0efb30",675:"ae9f91a8",733:"83105561",786:"06482edd",860:"03a8b7a5",873:"e6b09d52",898:"998ef92b"}[e]+"-1.236.0.min.js"),i.o=(e,t)=>Object.prototype.hasOwnProperty.call(e,t),e={},t="NRBA:",i.l=(r,n,o,a)=>{if(e[r])e[r].push(n);else{var s,c;if(void 0!==o)for(var u=document.getElementsByTagName("script"),d=0;d {s.onerror=s.onload=null,clearTimeout(h);var i=e[r];if(delete e[r],s.parentNode&&s.parentNode.removeChild(s),i&&i.forEach((e=>e(n))),t)return t(n)},h=setTimeout(l.bind(null,void 0,{type:"timeout",target:s}),12e4);s.onerror=l.bind(null,s.onerror),s.onload=l.bind(null,s.onload),c&&document.head.appendChild(s)}},i.r=e=>{"undefined"!=typeof Symbol&&Symbol.toStringTag&&Object.defineProperty(e,Symbol.toStringTag,{value:"Module"}),Object.defineProperty(e,"__esModule",{value:!0})},i.j=364,i.p="https://js-agent.newrelic.com/",(()=>{var e={364:0,953:0};i.f.j=(t,r)=>{var n=i.o(e,t)?e[t]:void 0;if(0!==n)if(n)r.push(n[2]);else{var o=new Promise(((r,i)=>n=e[t]=[r,i]));r.push(n[2]=o);var a=i.p+i.u(t),s=new Error;i.l(a,(r=>{if(i.o(e,t)&&(0!==(n=e[t])&&(e[t]=void 0),n)){var o=r&&("load"===r.type?"missing":r.type),a=r&&r.target&&r.target.src;s.message="Loading chunk "+t+" failed.\n("+o+": "+a+")",s.name="ChunkLoadError",s.type=o,s.request=a,n[1](s)}}),"chunk-"+t,t)}};var t=(t,r)=>{var n,o,[a,s,c]=r,u=0;if(a.some((t=>0!==e[t]))){for(n in s)i.o(s,n)&&(i.m[n]=s[n]);if(c)c(i)}for(t&&t(r);u {i.r(o);var e=i(3325),t=i(5763);const r=Object.values(e.D);function n(e){const n={};return r.forEach((r=>{n[r]=function(e,r){return!1!==(0,t.Mt)(r,"".concat(e,".enabled"))}(r,e)})),n}var a=i(9144);var s=i(5546),c=i(385),u=i(8e3),d=i(5938),f=i(3960),l=i(50);class h extends d.W{constructor(e,t,r){let n=!(arguments.length>3&&void 0!==arguments[3])||arguments[3];super(e,t,r),this.auto=n,this.abortHandler,this.featAggregate,this.onAggregateImported,n&&(0,u.R)(e,r)}importAggregator(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:{};if(this.featAggregate||!this.auto)return;const r=c.il&&!0===(0,t.Mt)(this.agentIdentifier,"privacy.cookies_enabled");let n;this.onAggregateImported=new Promise((e=>{n=e}));const o=async()=>{let t;try{if(r){const{setupAgentSession:e}=await Promise.all([i.e(860),i.e(242)]).then(i.bind(i,3228));t=e(this.agentIdentifier)}}catch(e){(0,l.Z)("A problem occurred when starting up session manager. This page will not start or extend any session.",e)}try{if(!this.shouldImportAgg(this.featureName,t))return void(0,u.L)(this.agentIdentifier,this.featureName);const{lazyFeatureLoader:r}=await i.e(412).then(i.bind(i,8582)),{Aggregate:o}=await r(this.featureName,"aggregate");this.featAggregate=new o(this.agentIdentifier,this.aggregator,e),n(!0)}catch(e){(0,l.Z)("Downloading and initializing ".concat(this.featureName," failed..."),e),this.abortHandler?.(),n(!1)}};c.il?(0,f.b)((()=>o()),!0):o()}shouldImportAgg(r,n){return r!==e.D.sessionReplay||!1!==(0,t.Mt)(this.agentIdentifier,"session_trace.enabled")&&(!!n?.isNew||!!n?.state.sessionReplay)}}var g=i(7633),p=i(7894);class m extends h{static featureName=g.t9;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];if(super(r,n,g.t9,i),("undefined"==typeof PerformanceNavigationTiming||c.Tt)&&"undefined"!=typeof PerformanceTiming){const n=(0,t.OP)(r);n[g.Dz]=Math.max(Date.now()-n.offset,0),(0,f.K)((()=>n[g.qw]=Math.max((0,p.z)()-n[g.Dz],0))),(0,f.b)((()=>{const t=(0,p.z)();n[g.OJ]=Math.max(t-n[g.Dz],0),(0,s.p)("timing",["load",t],void 0,e.D.pageViewTiming,this.ee)}))}this.importAggregator()}}var v=i(1117),b=i(1284);class y extends v.w{constructor(e){super(e),this.aggregatedData={}}store(e,t,r,n,i){var o=this.getBucket(e,t,r,i);return o.metrics=function(e,t){t||(t={count:0});return t.count+=1,(0,b.D)(e,(function(e,r){t[e]=w(r,t[e])})),t}(n,o.metrics),o}merge(e,t,r,n,i){var o=this.getBucket(e,t,n,i);if(o.metrics){var a=o.metrics;a.count+=r.count,(0,b.D)(r,(function(e,t){if("count"!==e){var n=a[e],i=r[e];i&&!i.c?a[e]=w(i.t,n):a[e]=function(e,t){if(!t)return e;t.c||(t=x(t.t));return t.min=Math.min(e.min,t.min),t.max=Math.max(e.max,t.max),t.t+=e.t,t.sos+=e.sos,t.c+=e.c,t}(i,a[e])}}))}else o.metrics=r}storeMetric(e,t,r,n){var i=this.getBucket(e,t,r);return i.stats=w(n,i.stats),i}getBucket(e,t,r,n){this.aggregatedData[e]||(this.aggregatedData[e]={});var i=this.aggregatedData[e][t];return i||(i=this.aggregatedData[e][t]={params:r||{}},n&&(i.custom=n)),i}get(e,t){return t?this.aggregatedData[e]&&this.aggregatedData[e][t]:this.aggregatedData[e]}take(e){for(var t={},r="",n=!1,i=0;i t.max&&(t.max=e),e 2&&void 0!==arguments[2])||arguments[2];super(e,r,j.t,n),c.il&&((0,t.OP)(e).initHidden=Boolean("hidden"===document.visibilityState),(0,N.N)((()=>(0,s.p)("docHidden",[(0,p.z)()],void 0,j.t,this.ee)),!0),(0,O.bP)("pagehide",(()=>(0,s.p)("winPagehide",[(0,p.z)()],void 0,j.t,this.ee))),this.importAggregator())}}var P=i(3081);class C extends h{static featureName=P.t9;constructor(e,t){let r=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(e,t,P.t9,r),this.importAggregator()}}var R,I=i(2210),k=i(1214),H=i(2177),L={};try{R=localStorage.getItem("__nr_flags").split(","),console&&"function"==typeof console.log&&(L.console=!0,-1!==R.indexOf("dev")&&(L.dev=!0),-1!==R.indexOf("nr_dev")&&(L.nrDev=!0))}catch(e){}function z(e){try{L.console&&z(e)}catch(e){}}L.nrDev&&H.ee.on("internal-error",(function(e){z(e.stack)})),L.dev&&H.ee.on("fn-err",(function(e,t,r){z(r.stack)})),L.dev&&(z("NR AGENT IN DEVELOPMENT MODE"),z("flags: "+(0,b.D)(L,(function(e,t){return e})).join(", ")));var M=i(6660);class B extends h{static featureName=M.t;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(r,n,M.t,i),this.skipNext=0;try{this.removeOnAbort=new AbortController}catch(e){}const o=this;o.ee.on("fn-start",(function(e,t,r){o.abortHandler&&(o.skipNext+=1)})),o.ee.on("fn-err",(function(t,r,n){o.abortHandler&&!n[M.A]&&((0,I.X)(n,M.A,(function(){return!0})),this.thrown=!0,(0,s.p)("err",[n,(0,p.z)()],void 0,e.D.jserrors,o.ee))})),o.ee.on("fn-end",(function(){o.abortHandler&&!this.thrown&&o.skipNext>0&&(o.skipNext-=1)})),o.ee.on("internal-error",(function(t){(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,o.ee)})),this.origOnerror=c._A.onerror,c._A.onerror=this.onerrorHandler.bind(this),c._A.addEventListener("unhandledrejection",(t=>{const r=function(e){let t="Unhandled Promise Rejection: ";if(e instanceof Error)try{return e.message=t+e.message,e}catch(t){return e}if(void 0===e)return new Error(t);try{return new Error(t+(0,D.P)(e))}catch(e){return new Error(t)}}(t.reason);(0,s.p)("err",[r,(0,p.z)(),!1,{unhandledPromiseRejection:1}],void 0,e.D.jserrors,this.ee)}),(0,O.m$)(!1,this.removeOnAbort?.signal)),(0,k.gy)(this.ee),(0,k.BV)(this.ee),(0,k.em)(this.ee),(0,t.OP)(r).xhrWrappable&&(0,k.Kf)(this.ee),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}onerrorHandler(t,r,n,i,o){"function"==typeof this.origOnerror&&this.origOnerror(...arguments);try{this.skipNext?this.skipNext-=1:(0,s.p)("err",[o||new F(t,r,n),(0,p.z)()],void 0,e.D.jserrors,this.ee)}catch(t){try{(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,this.ee)}catch(e){}}return!1}}function F(e,t,r){this.message=e||"Uncaught error with no additional information",this.sourceURL=t,this.line=r}let U=1;const q="nr@id";function G(e){const t=typeof e;return!e||"object"!==t&&"function"!==t?-1:e===c._A?0:(0,I.X)(e,q,(function(){return U++}))}function V(e){if("string"==typeof e&&e.length)return e.length;if("object"==typeof e){if("undefined"!=typeof ArrayBuffer&&e instanceof ArrayBuffer&&e.byteLength)return e.byteLength;if("undefined"!=typeof Blob&&e instanceof Blob&&e.size)return e.size;if(!("undefined"!=typeof FormData&&e instanceof FormData))try{return(0,D.P)(e).length}catch(e){return}}}var X=i(7243);class W{constructor(e){this.agentIdentifier=e,this.generateTracePayload=this.generateTracePayload.bind(this),this.shouldGenerateTrace=this.shouldGenerateTrace.bind(this)}generateTracePayload(e){if(!this.shouldGenerateTrace(e))return null;var r=(0,t.DL)(this.agentIdentifier);if(!r)return null;var n=(r.accountID||"").toString()||null,i=(r.agentID||"").toString()||null,o=(r.trustKey||"").toString()||null;if(!n||!i)return null;var a=(0,_.M)(),s=(0,_.Ht)(),c=Date.now(),u={spanId:a,traceId:s,timestamp:c};return(e.sameOrigin||this.isAllowedOrigin(e)&&this.useTraceContextHeadersForCors())&&(u.traceContextParentHeader=this.generateTraceContextParentHeader(a,s),u.traceContextStateHeader=this.generateTraceContextStateHeader(a,c,n,i,o)),(e.sameOrigin&&!this.excludeNewrelicHeader()||!e.sameOrigin&&this.isAllowedOrigin(e)&&this.useNewrelicHeaderForCors())&&(u.newrelicHeader=this.generateTraceHeader(a,s,c,n,i,o)),u}generateTraceContextParentHeader(e,t){return"00-"+t+"-"+e+"-01"}generateTraceContextStateHeader(e,t,r,n,i){return i+"@nr=0-1-"+r+"-"+n+"-"+e+"----"+t}generateTraceHeader(e,t,r,n,i,o){if(!("function"==typeof c._A?.btoa))return null;var a={v:[0,1],d:{ty:"Browser",ac:n,ap:i,id:e,tr:t,ti:r}};return o&&n!==o&&(a.d.tk=o),btoa((0,D.P)(a))}shouldGenerateTrace(e){return this.isDtEnabled()&&this.isAllowedOrigin(e)}isAllowedOrigin(e){var r=!1,n={};if((0,t.Mt)(this.agentIdentifier,"distributed_tracing")&&(n=(0,t.P_)(this.agentIdentifier).distributed_tracing),e.sameOrigin)r=!0;else if(n.allowed_origins instanceof Array)for(var i=0;i 2&&void 0!==arguments[2])||arguments[2];super(r,n,Z.t,i),(0,t.OP)(r).xhrWrappable&&(this.dt=new W(r),this.handler=(e,t,r,n)=>(0,s.p)(e,t,r,n,this.ee),(0,k.u5)(this.ee),(0,k.Kf)(this.ee),function(r,n,i,o){function a(e){var t=this;t.totalCbs=0,t.called=0,t.cbTime=0,t.end=E,t.ended=!1,t.xhrGuids={},t.lastSize=null,t.loadCaptureCalled=!1,t.params=this.params||{},t.metrics=this.metrics||{},e.addEventListener("load",(function(r){_(t,e)}),(0,O.m$)(!1)),c.IF||e.addEventListener("progress",(function(e){t.lastSize=e.loaded}),(0,O.m$)(!1))}function s(e){this.params={method:e[0]},T(this,e[1]),this.metrics={}}function u(e,n){var i=(0,t.DL)(r);i.xpid&&this.sameOrigin&&n.setRequestHeader("X-NewRelic-ID",i.xpid);var a=o.generateTracePayload(this.parsedOrigin);if(a){var s=!1;a.newrelicHeader&&(n.setRequestHeader("newrelic",a.newrelicHeader),s=!0),a.traceContextParentHeader&&(n.setRequestHeader("traceparent",a.traceContextParentHeader),a.traceContextStateHeader&&n.setRequestHeader("tracestate",a.traceContextStateHeader),s=!0),s&&(this.dt=a)}}function d(e,t){var r=this.metrics,i=e[0],o=this;if(r&&i){var a=V(i);a&&(r.txSize=a)}this.startTime=(0,p.z)(),this.listener=function(e){try{"abort"!==e.type||o.loadCaptureCalled||(o.params.aborted=!0),("load"!==e.type||o.called===o.totalCbs&&(o.onloadCalled||"function"!=typeof t.onload)&&"function"==typeof o.end)&&o.end(t)}catch(e){try{n.emit("internal-error",[e])}catch(e){}}};for(var s=0;s 1?e[1]=i:e.push(i)}else e[0]&&e[0].headers&&s(e[0].headers,n)&&(this.dt=n);function s(e,t){var r=!1;return t.newrelicHeader&&(e.set("newrelic",t.newrelicHeader),r=!0),t.traceContextParentHeader&&(e.set("traceparent",t.traceContextParentHeader),t.traceContextStateHeader&&e.set("tracestate",t.traceContextStateHeader),r=!0),r}}function x(e,t){this.params={},this.metrics={},this.startTime=(0,p.z)(),this.dt=t,e.length>=1&&(this.target=e[0]),e.length>=2&&(this.opts=e[1]);var r,n=this.opts||{},i=this.target;"string"==typeof i?r=i:"object"==typeof i&&i instanceof Y?r=i.url:c._A?.URL&&"object"==typeof i&&i instanceof URL&&(r=i.href),T(this,r);var o=(""+(i&&i instanceof Y&&i.method||n.method||"GET")).toUpperCase();this.params.method=o,this.txSize=V(n.body)||0}function A(t,r){var n;this.endTime=(0,p.z)(),this.params||(this.params={}),this.params.status=r?r.status:0,"string"==typeof this.rxSize&&this.rxSize.length>0&&(n=+this.rxSize);var o={txSize:this.txSize,rxSize:n,duration:(0,p.z)()-this.startTime};i("xhr",[this.params,o,this.startTime,this.endTime,"fetch"],this,e.D.ajax)}function E(t){var r=this.params,n=this.metrics;if(!this.ended){this.ended=!0;for(var o=0;o 2&&void 0!==arguments[2])||arguments[2];super(e,t,we.t,r),this.importAggregator()}}new class{constructor(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:(0,_.ky)(16);c._A?(this.agentIdentifier=t,this.sharedAggregator=new y({agentIdentifier:this.agentIdentifier}),this.features={},this.desiredFeatures=new Set(e.features||[]),this.desiredFeatures.add(m),Object.assign(this,(0,a.j)(this.agentIdentifier,e,e.loaderType||"agent")),this.start()):(0,l.Z)("Failed to initial the agent. Could not determine the runtime environment.")}get config(){return{info:(0,t.C5)(this.agentIdentifier),init:(0,t.P_)(this.agentIdentifier),loader_config:(0,t.DL)(this.agentIdentifier),runtime:(0,t.OP)(this.agentIdentifier)}}start(){const t="features";try{const r=n(this.agentIdentifier),i=[...this.desiredFeatures];i.sort(((t,r)=>e.p[t.featureName]-e.p[r.featureName])),i.forEach((t=>{if(r[t.featureName]||t.featureName===e.D.pageViewEvent){const n=function(t){switch(t){case e.D.ajax:return[e.D.jserrors];case e.D.sessionTrace:return[e.D.ajax,e.D.pageViewEvent];case e.D.sessionReplay:return[e.D.sessionTrace];case e.D.pageViewTiming:return[e.D.pageViewEvent];default:return[]}}(t.featureName);n.every((e=>r[e]))||(0,l.Z)("".concat(t.featureName," is enabled but one or more dependent features has been disabled (").concat((0,D.P)(n),"). This may cause unintended consequences or missing data...")),this.features[t.featureName]=new t(this.agentIdentifier,this.sharedAggregator)}})),(0,T.Qy)(this.agentIdentifier,this.features,t)}catch(e){(0,l.Z)("Failed to initialize all enabled instrument classes (agent aborted) -",e);for(const e in this.features)this.features[e].abortHandler?.();const r=(0,T.fP)();return delete r.initializedAgents[this.agentIdentifier]?.api,delete r.initializedAgents[this.agentIdentifier]?.[t],delete this.sharedAggregator,r.ee?.abort(),delete r.ee?.get(this.agentIdentifier),!1}}}({features:[J,m,S,class extends h{static featureName=oe;constructor(t,r){if(super(t,r,oe,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;const n=this.ee;let i;(0,k.QU)(n),this.eventsEE=(0,k.em)(n),this.eventsEE.on(se,(function(e,t){this.bstStart=(0,p.z)()})),this.eventsEE.on(ae,(function(t,r){(0,s.p)("bst",[t[0],r,this.bstStart,(0,p.z)()],void 0,e.D.sessionTrace,n)})),n.on(ce+ne,(function(e){this.time=(0,p.z)(),this.startPath=location.pathname+location.hash})),n.on(ce+ie,(function(t){(0,s.p)("bstHist",[location.pathname+location.hash,this.startPath,this.time],void 0,e.D.sessionTrace,n)}));try{i=new PerformanceObserver((t=>{const r=t.getEntries();(0,s.p)(te,[r],void 0,e.D.sessionTrace,n)})),i.observe({type:re,buffered:!0})}catch(e){}this.importAggregator({resourceObserver:i})}},C,xe,B,class extends h{static featureName=de;constructor(e,r){if(super(e,r,de,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;if(!(0,t.OP)(e).xhrWrappable)return;try{this.removeOnAbort=new AbortController}catch(e){}let n,i=0;const o=this.ee.get("tracer"),a=(0,k._L)(this.ee),s=(0,k.Lg)(this.ee),u=(0,k.BV)(this.ee),d=(0,k.Kf)(this.ee),f=this.ee.get("events"),l=(0,k.u5)(this.ee),h=(0,k.QU)(this.ee),g=(0,k.Gm)(this.ee);function m(e,t){h.emit("newURL",[""+window.location,t])}function v(){i++,n=window.location.hash,this[ve]=(0,p.z)()}function b(){i--,window.location.hash!==n&&m(0,!0);var e=(0,p.z)();this[pe]=~~this[pe]+e-this[ve],this[ye]=e}function y(e,t){e.on(t,(function(){this[t]=(0,p.z)()}))}this.ee.on(ve,v),s.on(be,v),a.on(be,v),this.ee.on(ye,b),s.on(ge,b),a.on(ge,b),this.ee.buffer([ve,ye,"xhr-resolved"],this.featureName),f.buffer([ve],this.featureName),u.buffer(["setTimeout"+le,"clearTimeout"+fe,ve],this.featureName),d.buffer([ve,"new-xhr","send-xhr"+fe],this.featureName),l.buffer([me+fe,me+"-done",me+he+fe,me+he+le],this.featureName),h.buffer(["newURL"],this.featureName),g.buffer([ve],this.featureName),s.buffer(["propagate",be,ge,"executor-err","resolve"+fe],this.featureName),o.buffer([ve,"no-"+ve],this.featureName),a.buffer(["new-jsonp","cb-start","jsonp-error","jsonp-end"],this.featureName),y(l,me+fe),y(l,me+"-done"),y(a,"new-jsonp"),y(a,"jsonp-end"),y(a,"cb-start"),h.on("pushState-end",m),h.on("replaceState-end",m),window.addEventListener("hashchange",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("load",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("popstate",(function(){m(0,i>1)}),(0,O.m$)(!0,this.removeOnAbort?.signal)),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}}],loaderType:"spa"})})(),window.NRBA=o})(); window.jQuery || document.write(' ') CKEDITOR_BASEPATH='https://f1000research.com/js/vendor/ckeditor/' window.reactTheme = 'research'; window.MathJax = { CommonHTML: { linebreaks: { automatic: true } }, 'HTML-CSS': { linebreaks: { automatic: true } }, SVG: { linebreaks: { automatic: true } }, AuthorInit: function() { MathJax.Hub.Register.MessageHook('End Process', function () { let timeout = false; // holder for timeout id const delay = 250; // delay after event is "complete" to run callback const reflowMath = function() { const dispFormulas = document.querySelectorAll('.disp-formula.panel'); if (!dispFormulas) { return; } for (const dispFormula of dispFormulas) { const child = dispFormula.querySelector('.MathJax_Preview').nextSibling.firstChild; const isMultiline = MathJax.Hub.getAllJax(dispFormula)[0].root.isMultiline; if (dispFormula.offsetWidth < child.offsetWidth || isMultiline) { MathJax.Hub.Queue(['Rerender', MathJax.Hub, dispFormula]); } } }; window.addEventListener('resize', function() { clearTimeout(timeout); // clear the timeout timeout = setTimeout(reflowMath, delay); // start timing for event "completion" }); }); }, }; if (window.location.hash == '#_=_'){ window.location = window.location.href.split('#')[0] } !function(f,b,e,v,n,t,s){if(f.fbq)return;n=f.fbq=function() {n.callMethod? n.callMethod.apply(n,arguments):n.queue.push(arguments)} ;if(!f._fbq)f._fbq=n; n.push=n;n.loaded=!0;n.version='2.0';n.queue=[];t=b.createElement(e);t.async=!0; t.src=v;s=b.getElementsByTagName(e)[0];s.parentNode.insertBefore(t,s)}(window, document,'script','https://connect.facebook.net/en_US/fbevents.js'); fbq('init', '1641728616063202'); fbq('track', "PixelInitialized", {}); (function(h,o,t,j,a,r){ h.hj=h.hj||function(){(h.hj.q=h.hj.q||[]).push(arguments)}; h._hjSettings={hjid:2318163,hjsv:6}; a=o.getElementsByTagName('head')[0]; r=o.createElement('script');r.async=1; r.src=t+h._hjSettings.hjid+j+h._hjSettings.hjsv; a.appendChild(r); })(window,document,'https://static.hotjar.com/c/hotjar-','.js?sv='); search file_upload Submit your research search menu close search Browse Gateways & Collections How to Publish Submit your Research My Submissions Article Guidelines Article Guidelines (New Versions) Open Data, Software and Code Guidelines Open Data and Accessible Source Materials Guidelines (HSS) Open Data, Software and Code Guidelines (PSE) Prepublication Checks Production Process Posters and Slides Guidelines Document Guidelines Article Processing Charges Peer Review Finding Article Reviewers About How it Works For Reviewers Our Advisors Policies Glossary FAQs For Developers Newsroom Contact My Research Submissions Content and Tracking Alerts My Details Sign In file_upload Submit your research <input type="hidden" id="meta-article-title" value=" Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study" /> { "@context": "https://schema.org", "@type": "ScholarlyArticle", "mainEntityOfPage": { "@type": "WebPage", "@id": "https://f1000research.com/articles/13-639" }, "headline": "Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1...", "datePublished": "2024-06-17T15:05:40", "dateModified": "2024-10-22T17:28:53", "author": [ { "@type": "Person", "name": "Manjunath Shetty" }, { "@type": "Person", "name": "Smita Shenoy" }, { "@type": "Person", "name": "Arul Amuthan" }, { "@type": "Person", "name": "Vasudha Devi" }, { "@type": "Person", "name": "Nitesh Kumar" }, { "@type": "Person", "name": "Amruth Kiran" }, { "@type": "Person", "name": "Ganesh Shenoy" }, { "@type": "Person", "name": "Diya Rajasekhar Chinta" }, { "@type": "Person", "name": "Shama Prasada K" }, { "@type": "Person", "name": "Akshatha Shetty" }, { "@type": "Person", "name": "Mohandas Rao K G" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": " Background Hepatocellular carcinoma (HCC) is an aggressive malignancy with poor clinical outcomes. Hence cost-effective drugs with fewer side effects as a standard supportive therapy might yield substantial advantages in efficacy and safety. Kadukkai maathirai (KM) is being used as a supplement in hepatocellular carcinoma. We evaluated whether KM has any preventive action on cancer progression in diethyl nitrosamine (DEN) - induced HCC in rats. Methods DEN was injected to produce HCC in rats, which was confirmed after 16 weeks. All the rats were orally administered KM for 4 weeks. Hepatoprotective potential (serum AST, ALT, ALP, Bilirubin) and anticancer efficacy (body weight, nodule count, tumor progression by histopathology, expression of GSTM1 by Liquid chromatography-mass spectrometry (LC-MS), and In-silico analysis of phytoconstituents against β catenin and LRP analysis were evaluated. Results KM prevented cancer progression against DEN-induced HCC by an increase in GSTM1, a phase II detoxifying enzyme. It significantly reversed altered nodule count, relative liver weight, body weight, and histopathological features of HCC. In silico analysis of phytoconstituents of KM showed that they modulate the intracellular transcription process by inhibiting the armadillo repeat region of β catenin. Conclusions Our results elucidate the potential of KM as a supplement in HCC by reducing nodule count, protecting the liver from further damage, GSTM1 expression, and inhibiting armadillo repeat region of β catenin. " } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/13-639/v2", "name": "Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer..." } } ] } Home Browse Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Shetty M, Shenoy S, Amuthan A et al. Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.12688/f1000research.145961.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Revised Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] Manjunath Shetty https://orcid.org/0000-0002-3106-3683 1,2 , Smita Shenoy https://orcid.org/0000-0002-7578-1855 3 , Arul Amuthan 2 , [...] Vasudha Devi 3 , Nitesh Kumar https://orcid.org/0000-0002-4929-3954 4 , Amruth Kiran 2 , Ganesh Shenoy 2 , Diya Rajasekhar Chinta 5 , Shama Prasada K https://orcid.org/0000-0002-4158-3893 6 , Akshatha Shetty 7 , Mohandas Rao K G 8 Manjunath Shetty https://orcid.org/0000-0002-3106-3683 1,2 , Smita Shenoy https://orcid.org/0000-0002-7578-1855 3 , [...] Arul Amuthan 2 , Vasudha Devi 3 , Nitesh Kumar https://orcid.org/0000-0002-4929-3954 4 , Amruth Kiran 2 , Ganesh Shenoy 2 , Diya Rajasekhar Chinta 5 , Shama Prasada K https://orcid.org/0000-0002-4158-3893 6 , Akshatha Shetty 7 , Mohandas Rao K G 8 PUBLISHED 22 Oct 2024 Author details Author details 1 Centre Of Excellence, Ocular Nanoscience, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 2 Division of Pharmacology, Department of Basic Medical Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 3 Department of Pharmacology, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 4 Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, Vaishali, Bihar, 844102, India 5 Department of Pharmacology, Manipal University College Malaysia, Bukit Baru, Melaka, 75150, Malaysia 6 Department of Cell and Molecular Biology, School of Life Sciences, Manipal Academy of Higher Education, Manipal, Manipal, Karnataka, 576104, India 7 Department of Research and Development, Muniyal Institute of Ayurveda and Medical Sciences, Manipal, Manipal, Karnataka, 576104, India 8 Division of Anatomy, Department of Basic Medical Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India Manjunath Shetty Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Smita Shenoy Roles: Project Administration, Supervision, Validation, Writing – Review & Editing Arul Amuthan Roles: Conceptualization, Methodology, Supervision Vasudha Devi Roles: Supervision Nitesh Kumar Roles: Methodology, Supervision, Validation, Visualization Amruth Kiran Roles: Resources, Software, Visualization, Writing – Review & Editing Ganesh Shenoy Roles: Investigation, Visualization, Writing – Review & Editing Diya Rajasekhar Chinta Roles: Software, Validation, Visualization, Writing – Review & Editing Shama Prasada K Roles: Software, Validation, Writing – Review & Editing Akshatha Shetty Roles: Validation, Visualization, Writing – Review & Editing Mohandas Rao K G Roles: Methodology, Resources, Validation, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Manipal Academy of Higher Education gateway. Abstract Background Hepatocellular carcinoma (HCC) is an aggressive malignancy with poor clinical outcomes. Hence cost-effective drugs with fewer side effects as a standard supportive therapy might yield substantial advantages in efficacy and safety. Kadukkai maathirai (KM) is being used as a supplement in hepatocellular carcinoma. We evaluated whether KM has any preventive action on cancer progression in diethyl nitrosamine (DEN) - induced HCC in rats. Methods DEN was injected to produce HCC in rats, which was confirmed after 16 weeks. All the rats were orally administered KM for 4 weeks. Hepatoprotective potential (serum AST, ALT, ALP, Bilirubin) and anticancer efficacy (body weight, nodule count, tumor progression by histopathology, expression of GSTM1 by Liquid chromatography-mass spectrometry (LC-MS), and In-silico analysis of phytoconstituents against β catenin and LRP analysis were evaluated. Results KM prevented cancer progression against DEN-induced HCC by an increase in GSTM1, a phase II detoxifying enzyme. It significantly reversed altered nodule count, relative liver weight, body weight, and histopathological features of HCC. In silico analysis of phytoconstituents of KM showed that they modulate the intracellular transcription process by inhibiting the armadillo repeat region of β catenin. Conclusions Our results elucidate the potential of KM as a supplement in HCC by reducing nodule count, protecting the liver from further damage, GSTM1 expression, and inhibiting armadillo repeat region of β catenin. READ ALL READ LESS Keywords β catenin, Glutathione-S transferase mu 1, Siddha, herbal, Diethyl nitrosamine Corresponding Author(s) Smita Shenoy ( [email protected] ) Close Corresponding author: Smita Shenoy Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2024 Shetty M et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Shetty M, Shenoy S, Amuthan A et al. Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.12688/f1000research.145961.2 ) First published: 17 Jun 2024, 13 :639 ( https://doi.org/10.12688/f1000research.145961.1 ) Latest published: 22 Oct 2024, 13 :639 ( https://doi.org/10.12688/f1000research.145961.2 ) Revised Amendments from Version 1 1.Author affiliations: KG Mohandas Rao affiliation has been changed to correct an error. 2.Discussion: Corrections have been made to explain the findings with reason wherever possible and improve the grammar. The reason for all findings cannot be explicitly stated as this requires detailed mechanistic study which is the future scope and is accordingly mentioned in the conclusion. Study findings have been linked to earlier reports in literature. Discussion has been organized to improve the flow. One reference is deleted and one added as follows- Reference 63 is deleted. 64 renumber as 63. 65 renumber as 64. And new reference will be numbered as 65. 3. Conclusion: Heading is corrected as Conclusion (instead of Conclusions). Conclusion has been corrected to reflect the potential role of Kadukkai maathirai in therapy along with further scope of the study. 1.Author affiliations: KG Mohandas Rao affiliation has been changed to correct an error. 2.Discussion: Corrections have been made to explain the findings with reason wherever possible and improve the grammar. The reason for all findings cannot be explicitly stated as this requires detailed mechanistic study which is the future scope and is accordingly mentioned in the conclusion. Study findings have been linked to earlier reports in literature. Discussion has been organized to improve the flow. One reference is deleted and one added as follows- Reference 63 is deleted. 64 renumber as 63. 65 renumber as 64. And new reference will be numbered as 65. 3. Conclusion: Heading is corrected as Conclusion (instead of Conclusions). Conclusion has been corrected to reflect the potential role of Kadukkai maathirai in therapy along with further scope of the study. To read any peer review reports and author responses for this article, follow the "read" links in the Open Peer Review table. READ REVIEWER RESPONSES Introduction Hepatocellular carcinoma (HCC) is among the sixth most common and the second most lethal cancers. 1 It may occur as a sequel to chronic liver conditions such as alcohol and tobacco consumption, obesity, and infections like hepatitis B, hepatitis C, and aflatoxins. 2 , 3 These factors lead to liver inflammation and fibrosis, followed by the destruction of the normal liver architecture. Chemotherapy, radiotherapy, radiofrequency ablation, hepatectomy, and transplantation are the management plans of HCC. 4 However, the survival rate is low, suggesting the need for improved therapy. 5 Chemotherapeutic agents used include sorafenib (MAP-kinase inhibitor), doxorubicin, 5-fluorouracil (5-FU), and cisplatin. 6 Being a multikinase inhibitor sorafenib, acts on vascular endothelial growth factor (VEGF) receptors, and prolongs survival so this drug is considered as a new reference standard for advanced HCC. However, it elicits adverse effects including skin toxicities, diarrhea, and hypertension, along with other general toxicities like anorexia, hair loss, weight loss, etc. 7 Compared to conventional modes of treatment, plant-based medicines have generally fewer side effects, are cost-effective, and well tolerated by patients. 8 Kadukkai maathirai (KM) is an Indian traditional medicine prescribed by Siddha practitioners to treat edema in severe anemia and ascites secondary to liver disease. KM consists of Terminalia chebula Retz. (Combretaceae ), Piper nigrum L. (Piperaceae) , Eclipta alba L. (Asteraceae), Citrus L. (Rutaceae) and ferrous sulfate. 9 Each of the constituents mentioned is known to have a hepatoprotective effect. 10 T. chebula consists of gallic acid and chebulic acid; gallic acid has anti-inflammatory and antioxidant properties, whereas chebulic acid has antioxidant with hepatoprotective properties. 11 P. nigrum inhibits lipid peroxidation and superoxide free radical generation due to its antioxidant effect exerted by phenols and flavonoids. 12 , 13 Coumestans, present in E. alba , stimulates the regeneration of hepatocytes and protect the liver. 14 Though Kadukkai maathirai is used for treating liver disorders by practitioners of Siddha medicine, yet its effect on hepatocellular carcinoma has not been explored. Diethylnitrosamine (DEN) is a known potential hepatocarcinogen that disrupts nuclear enzymes that help in the repair of DNA and replication. 14 In our previous study we demonstrated that the KM at the dose of 144 mg/kg treatment exhibited significant protection which was evident both by histology and biochemically against D-galactosamine-induced hepatic necrosis in rats. 15 Hepatic DEN metabolism produces O6-ethyl deoxyguanosine and O4- and O6-ethyl deoxythymidine, which are mutagenic and carcinogenic. 16 , 17 The effect of analyzed phytoconstituents of KM on low-density lipoprotein (LDL) receptor-related protein-6 (LRP6) and β-catenin signaling pathway, which play a role in hepatocellular carcinoma (HCC), was also studied in silico. Methods Chemicals and consumables Diethylnitrosamine 0.01% (Catalogue number - D0516) from TCI chemicals, Tokyo, Japan, assay kits for liver function test from Spain-based Spinreact, silymarin 50mg/kg body weight/day (Item Number – 46791) from Micro Labs Limited, India, Kadukkai maathirai (MMP14013) from India-based SKM Siddha and Ayurveda (GMP certified) Company (India) Ltd. Good laboratory-grade chemicals were used. Animals Ethical statement: Seven-week-old female Sprague Dawley rats (150-200 g), procured from the Central Animal Research Facilities (CARF) of Kasturba Medical College, Manipal Academy of Higher Education, Manipal were used in the study after getting approval from the Institutional Animal Ethics Committee approval (IAEC/KMC/19/2016 dated 16.03.2016). Guidelines given by the Committee for Control and Supervision of Experiments on Animals (CCSEA), Government of India, New Delhi for the use of laboratory animals were followed for the maintenance of animals. 18 All efforts were made to ameliorate suffering of animals. All the rats were observed daily to check their health and ensure continuous access to food and water. Bedding in the cages were changed daily to provide healthy living conditions. The location of the animal cage was not changed throughout the study. The rats were acclimatized and maintained at 27 ± 3°C, humidity of 60 ± 10%, and a 12 h light/dark cycle. No procedures were done which would cause sustained pain. Anesthetics and the method of euthanasia used was as per guidelines of CCSEA. Reporting Guidelines: The authors confirm that they followed and adhered to ARRIVE 2.0 Checklist for their study. ARRIVE CHECKLIST [Internet]. figshare; 2024 [cited 2024 Jun 7]. Available from: https://figshare.com/articles/dataset/ARRIVE_CHECKLIST/25910563/1. 76 Study design: The rats were randomised into 2 groups based on their body weight: normal control (n=6) and toxic control (n=30) rats. Induction of liver cancer: The number (n=6) of animals in each group was based on earlier study. 19 The groups were as follows: The normal control (group 1) consisted of a total of 6 rats and received drinking water for 16 weeks. The toxic control group (DEN) consisted of a total of 30 rats, which received DEN 0.01% for sixteen weeks through drinking water for induction of HCC. 20 At the end of 16 weeks, animals were divided into 5 groups (n=6/group) and received treatment orally as follows. Group 2- toxicant control and received gum acacia 2% 1 mL/kg orally for 4 weeks. 21 Group 3, 4, 5- test drug KM36, 72, 144 mg/kg body weight orally for 4 weeks respectively. 22 Group 6- received Silymarin (50 mg/kg body weight) orally for 4 weeks. 23 At the end of the study (total of 20 weeks) blood was drawn by retroorbital puncture. The animals of all groups were euthanised by administering pentobarbitone intraperitoneally in a dose of 200 mg/kg body weight, 24 then livers were dissected for histopathology and polyacrylamide gel electrophoresis. Parameters assessed • Parameters to evaluate hepatoprotective potential of KM After 4 weeks of KM treatment, under ketamine anesthesia (50 mg/kg i.p), 25 blood was withdrawn from all the rats from each group to estimate the serum AST, ALT, ALP, bilirubin, and total protein levels using kits (Aspen Laboratory). 26 • Parameters to evaluate whether KM has any anticancer effect Determination of liver weight and nodule count The liver of each animal was weighed and relative weight was calculated. The nodules on the liver were counted, and an intergroup comparison was made. Determination of body weight Weight was measured at the end of study by using a metis electronic weighing balance. The weight difference was calculated. Histopathological evaluation 10% formalin was used as the fixative. Paraffin-embedded wax blocks were prepared and the tissue section was prepared at 4μm thickness. The hepatic tissue was stained with hematoxylin-eosin and studied. 27 Regarding the animal group, the assessor was blinded. Polyacrylamide gel electrophoresis for the separation of proteins and mass spectrometry for the identification of proteins. Sample Preparation and Electrospray-ionisation Quadrapole time of fight mass (ESI/QTOF) liquid chromatography and mass spectrometry analysis (LC-MS). Tissue extract was prepared, and proteins were separated by polyacrylamide gel electrophoresis. LC-MS analysis was used to identify the protein that was differentially expressed in response to DEN administration. The expressed protein was identified by coomassie staining, separated from PAGE gel, and subjected to LC-MS analysis using MASCOT Ver 2.3. 28 In-silico analysis of phytoconstituents of KM against β catenin and LRP6 in HCC The binding mode and interaction of β catenin and LRP6-E3 with each of the derived list of phytoconstituents of Kadukkai maathirai was performed using AutoDock Vina software. 29 , 30 Grid boxes were set at two binding sites of the armadillo repeat region of β catenin and their XYZ coordinates include X: 107.640 Y: -29.892 Z: 1.991 Å and X: 102.640 Y: 9.850 Z: 29.741 Å respectively with a grid size of 40. Whereas the XYZ coordinates for LRP6-E3 were defined by establishing a grid box with the dimensions of X: -29.376 Y: 20.133 Z: -10.560 Å with a grid size of 40, The interactions of β catenin and LRP6 protein-ligand conformations, including hydrogen, and hydrophobic bonds along with their interacting residues were analyzed using BIOVIA Discovery Studio Visualizer. 31 The 2D structures of phytoconstituents present in KM were collected from the PubChem Compound database. A total of 19 structures were sketched, prepared, and saved in MDL mol format using ChemSketch 2019 2.2 32 and saved as PDBQT format. The 3D structure of β catenin (PDB ID: 1JPW) and LRP6 (PDB ID: 3S8V) were retrieved from Protein Data Bank (RCSB PDB), optimized, minimized their energy and saved as PDB format. Analysis of extract using Gas chromatography-mass spectroscopy (GC-MS) It was done with Shimadzu -QP2010. Column (30m length) used was RTX-5. Interface temperature 280 °C and ion source was 200 °C. Sample was injected (1 μL) for analysis and flow rate retained at 1 mL/minute. MS spectrum achieved was identified and compared with NIST library for confirmation of compounds. 33 Statistical analysis The data of control, test drug control, and experimental groups were presented in the form of mean ± SD, and one-way ANOVA and post hoc Tukey's test (SPSS version 16) were performed. Statistical significance was considered at p<0.05. Results Hepatoprotective potential of KM Biochemical estimation in serum Serum AST, ALP, ALT, and TBIL in DEN alone treated animals increased significantly in comparison to normal control indicating liver damage ( Table 1 ). KM 36 mg/kg and KM72 mg/kg in DEN-treated groups showed a fall in the AST, ALT, ALP, and TBIL levels compared to animals that received DEN alone but it was not statistically significant. Other parameters were not affected significantly ( Table 1 ). Table 1. Effect of KM on liver function in DEN-induced HCC model in rats. Groups Serum AST (U/L) Serum ALT (U/L) Serum ALP (U/L) Serum TBIL (mg/dL) Serum Albumin (g/dL) Normal control 59.0±7.5 35.4±8.7 120.8±20.0 0.03±0.01 1.44±0.3 DEN 145.5±24.1 * 99.0±14.8 * 274.3±69.1 * 0.126±0.08 * 0.9±0.1 DEN+KM36 mg/kg 107.7±10.4 73.9±8.7 243.6±14.7 0.036±0.00 1.3±0.1 DEN+KM72 mg/kg 131.0±26.4 93.8±32.6 283.6±42.5 0.068±0.06 1.3±0.4 DEN+KM144 mg/kg 85.5±13.2 # , € , ¥ 71.1±10.0 257.25±26.2 0.035±0.02 0.9±0.1 DEN+Silymarin 136.2±37.1 83.15±18.8 280.75±82.9 0.099±0.02 0.9±0.1 * p<0.05 versus control. # p<0.05 versus DEN. € p<0.05 versus DEN+KM72 mg/kg. ¥ p<0.05 versus DEN+Silymarin, (Values - Mean ±SD). Bodyweight/Organ weight/Nodule count: Body weight decreased significantly and relative liver weight increased in the DEN-induced HCC group versus control. These were altered favorably(p<0.05) in DEN-treated rats who received KM 36, 72, 144 mg/kg, and silymarin as compared to DEN-alone treated rats ( Table 2 ). A decrease (p<0.05) in nodule count in rats treated with KM 36 mg/kg, KM 72 mg/kg, and silymarin was observed in comparison to DEN group ( Table 2 , Figure 1 ). Table 2. Effect of KM on body weight, relative weight of liver and nodule count in DEN-induced hepatocellular carcinoma model in rats. Groups Nodule count Relative weight of liver (g) Bodyweight(g) Normal control 0 0.02±0.002 257.5±8.8 DEN (Diethylnitrosamine) 107±4.0 * 0.09±0.017 * 165.3±4.1 * DEN+KM 36 mg/kg 72.0±8.1 # , € 0.03±0.005 # 222.6±9.5 # DEN+KM 72 mg/kg 84.4±11.8 # 0.04±0.004 # 200.2±3.7 # DEN+KM 144 mg/kg 97.5±2.6 0.04±0.005 # 212.7±16.8 # DEN+Silymarin 59.7±4.1 # , ¥ , € 0.04±0.007 # 200.0±8.4 # * p<0.05 versus control. # p < 0.05 versus DEN. € p< 0.05 versus DEN+KM144. ¥ p<0.05 versus DEN+KM72. Values: Mean±SD. Figure 1. Effect of KM on the gross (macroscopic) appearance of liver in DEN-induced HCC model in rats. Yellow arrows indicate nodules. 1A (Normal control): Normal morphology of liver with smooth-surfaced hepatic lobes in the control animal. 1B (DEN): DEN alone administered group showed highly enlarged nodules with several foci on the surface of the liver and many greyish-white nodules, indicative of HCC. 1C (DEN +KM 36 mg/kg), 1D (DEN +KM 72 mg/ kg), 1E (DEN+KM 144 mg/kg), 1F (DEN + silymarin 50 mg/kg) groups showed relatively less foci on the hepatic surface and very few nodules when compared to 1B (DEN). Histopathological evaluation of the liver Figure 2A , 2B (normal control): Showed normal liver structure, Figure 2C , 2D (DEN control) : Showed an invasive tumor involving irregular nodules divided by thin fibrous septae and composed of solid sheets and broad trabeculae of neoplastic hepatocytes with round to polygonal cells, irregular nuclei, prominent nucleoli, vesicular chromatin, moderate to abundant granular eosinophilic cytoplasm, mitotic figures, scattered with numerous variably dilated sinusoids, flattened endothelial cells and focal bile pigments. The presence of many areas of high vascularity with several groups of cut sections of blood vessels and destruction of the typical pattern of hepatic cords/hepatocyte arrangement is indicative of hepatocellular carcinoma. Figures 2E , 2F (DEN+KM36), Figures 2G , 2H (DEN+ KM72), Figures 2I , 2J (DEN+KM144 ): The structure of liver looked normal with hexagonal hepatic lobules, a central vein in its center. However, there were areas showing features of HCC like well-vascularized tumours with thick trabeculae, distinct acinar pattern, some areas of vascular invasion, and signs of mild cytoplasmic vacuolar degeneration. However, the severity of the features of HCC in this group was very mild when compared with that of DEN. Figures 2K , 2L (DEN+ Silymarin): There were areas of mild cytoplasmic vacuolar degeneration and high vascularity indicative of HCC. But it was less when compared with that of DEN group. In general, there were normal liver tissues in large areas compared to the areas with features of HCC. Figure 2. Effect of KM on histopathology of the liver in DEN-induced HCC model in rats; Qualitative assessment of ‘haematoxylin and eosin (H & E) stained liver tissue sections’ were observed under 10× and 40× magnifications; CV-Central vein, BV-Blood vessels, PT-Portal triad. Polyacrylamide Gel Electrophoresis for the separation of proteins and mass spectrometry for the identification of proteins Ladder: Lanes (1–4) - normal control and test drug control groups; Lane 5 - DEN alone, Lanes 6–9 - DEN for 16 weeks followed by treatment with KM 36, 72 and 144 mg/kg, and silymarin, respectively. The protein lysate was prepared and subjected to separation by PAGE ( Figure 3A ). DEN control group had a band in the range of 25-30 kda as analysed by Polyacrylamide Gel Electrophoresis (PAGE). In comparison to lanes 5 and 6, band thickness was more in lanes 7, 8, and 9; this was confirmed by mass spectrometry and identified as GSTM1 ( Figure 3B ). Figure 3. (A) Agarose gel electrophoresis of extraction from the liver of rats; (B) Mass spectrometry analysis of extraction from the liver of rats. In-silico analysis of interactions of phytoconstituents of KM with β catenin The molecular docking study revealed that nine phytoconstituents out of 19 made interactions with β catenin binding site that is residue number 219 to 335 (C-terminal helical region of Tcf-4interaction region) except D-allose which showed interactions between the residues 429 to 508 (extended region of Tcf-4 peptide interaction region) of β catenin. Their binding energies, number of interactions, and interacting residues of β catenin were tabulated in Table 3 . The binding energies of the phytoconstituents ranged from -7.8 to -5.2 kcal/mol. The lowest binding energy was recorded for hexanediamide whereas, 2H-Pyran-2-one, 5,6-dihydro documented the highest among the shortlisted constituents. The phytoconstituents with interacting residues are displayed in Figure 4A and 4B as follows. Octadecadienoicacid and propylene glycol monooleate made a maximum number of interactions (8 each) whereas, Hexanediamide made the lowest number of interactions- 2 ( Table 3 ). However, the constituents did not show any favorable interactions with the interaction site of LRP6-E3. Table 3. Shortlisted phytoconstituents and their interactions with β catenin. Phytoconstituents Binding energy No. of interactions Hydrogen bonds Hydrophobic bonds Hexanediamide -7.8 2 - 2(Phe253, lys252) Octadecadienoicacid -6.0 8 Lys335 7 (His260, Ile296, Lys292, Phe253) Propylene glycol monooleate -6.6 8 Arg376 7 (Tyr254, Phe253, Ile296, Lys292) Ethyl piperonylcyanoacetate -6.4 4 Asn290 3 (His219, Phe253, Lys292) Beta-sitosterol -7.3 5 Phe253 4(Ile296, Lys292, phe253) Benzene, 1-methyl-2-(1-methylethyl) -5.7 4 - 4 (Ile296, Lys292, Phe253) 2H-Pyran-2-one, 5,6-dihydro -5.2 2 - 2 (Lys292, Phe253) 3,5-dibromo-1H-pyrazole-4-carboxamide -5.5 4 - 4 (Ile296, Phe253) D-allose -6.5 4 4 (Lys435, His470) - Figure 4. Representation of the interactions made by the shortlisted phytoconstituents of KM with the binding site of β catenin. (Left: binding poses; Right: 2D images) (< 5 Å distance). Gas chromatography-mass spectroscopy (GC-MS) analysis of test drug KM ( Figure 5 ) ( Table 4 ) Figure 5. GC-MS analysis of the test drug Kadukkai maathirai (KM). Table 4. Gas chromatography-mass spectroscopy (GC-MS) analysis of the test dru g Kadukkai maathirai (KM). Sl. No R. time Compound name Mol. Formula Mol. Weight Percentage Structure 1 24.366 Piperine C 17 H 19 NO 3 285 66.26 2 11.173 D-Allose C 6 H 12 O 6 180.16 4.97 3 23.177 Propyleneglycol monooleate C 21 H 40 O 3 340 4.68 4 6.293 Levoglucosenone C 6 H 6 O 3 126 3.15 5 12.448 Caryophyllenyl alcohol C 15 H 26 O 222 2.57 6 8.740 Pentadecane C 15 H 32 212 2.06 7 6.964 Hexanediamide, N,N'-di-benzoyloxy- C 2 0H 2 0N 2 O 6 384 2.00 8 28.308 beta-Sitosterol C 29 H 50 O 414 1.72 9 22.366 Ethyl piperonylcyanoacetate C 13 H 13 NO 4 247 1.72 10 16.202 10,12-Octadecadienoic acid, 9-oxo- C 18 H 30 O 3 294 1.46 11 16.384 n-Hexadecanoic acid C 16 H 32 O 2 256 1.37 12 12.070 Dodecanoic acid C 12 H 24 O 2 200 1.18 13 18.077 cis-9-Hexadecenal C 16 H 30 O 238 1.13 14 7.629 1,4:3,6-Dianhydro-.alpha.-d-glucopyranose C 6 H 8 O 4 144.1 1.06 15 9.774 3a,7-Methano-3aH-cyclopentacyclooctene, 1,4,5,6,7,8,9,9a-octahydro-1,1,7-trimethyl-, [3 C 15 H 24 204 1.00 16 5.036 Benzene, 1-methyl-2-(1-methylethyl)- C 1 0H 14 134 0.94 17 4.131 3-Aminopyrazine 1-oxide C 4 H 5 N 3 O 111 0.94 18 5.228 2H-Pyran-2-one, 5,6-dihydro- C 5 H 6 O 2 98 0.90 19 3.328 Oxime-, methoxy-phenyl- C 8 H 9 NO 2 151 0.90 19 compounds were recognized by GC-MS from methanolic extract of KM. Piperine (66.26%) was the major constituent of KM. Others were D-Allose (4.97%), Propyleneglycol monoleate (4.68%), Levoglucosenone (3.15%), etc. Discussion N-nitroso compounds, as potential carcinogens, were first brought to the public attention as a cause of liver disease in 1937. In 1954, it was confirmed that one oral or parenteral dose of DEN (20-40 mg/kg) acts as a liver toxin, leading to severe liver necrosis in rodents. 34 , 35 In rodents, chronic parenteral and oral administration of DEN at large doses effectively induces liver tumour. 36 Pericentral foci of small dysplastic liver cells is induced by diethylnitrosamine (DEN), by ethylating DNA 37 , 38 leading to cirrhosis and multifocal HCC. 39 It predominantly targets the liver, where it is biologically transformed by CYP450-dependent processes that are most active in centrilobular hepatocytes. 40 The main metabolising pathways of DEN in rodents and humans are very similar. 41 which results in reactive species responsible for the development of methylated compounds, leading to carcinogenesis. Previous studies have shown that in DEN-treated group, there was distorted liver architecture with swollen cells, hyperplasia, and dysplasia, with infiltration of inflammatory cells, indicating carcinogenesis. 42 – 44 In the present study, similar histopathological changes occurred in DEN-treated groups. DEN-induced hepatic injury leads to instability of metabolism by the liver with concomitant variations in the activities of enzymes. 45 An increase in the level of transaminases indicates liver damage. Liver toxicity leads to the breakdown of the cell membrane and further spillage of the aminotransferases from the cytoplasm into the serum, thus increasing the serum aminotransferase levels. 46 Aminotransferases are known as diagnostic and reliable markers of liver damage. In our study, the level of liver enzymes and bilirubin were increased significantly reflecting liver damage in DEN alone treated rats. In rats which received DEN with KM, the enzyme levels were reduced when compared to DEN alone treated rats, though it was not statistically significant. Probably, increasing the duration of treatment (> 4 weeks) with KM may have produced further improvement in liver functions. Weight loss is an important symptom of DEN-induced cancer. DEN causes alterations which is reflected as worsening of liver function during pathogenesis of liver fibrosis. 47 Damaged to the liver results in loss of appetite. Administration of KM in varying doses following induction of HCC by DEN prevented body weight loss which could be due to its protective action against liver damage. In this study, the relative liver weights of DEN-treated rats were significantly high compared to normal which has been commonly observed and reported in liver cancer. 48 Treatment with KM prevented an increase in liver weight. DEN gets metabolized by microsomal enzymes in a sequence of intermediate responses leading to development of mutagenic metabolites. The free radicals generated in the process trigger cell proliferation, appearance of liver nodules and a rise in the weight of the liver. 49 Low levels of ROS do not cause any damage to the cell process, but high ROS level damages proteins, lipids and DNA by nonspecific process. 50 Glutathione-S transferase mu 1 (GSTM1) is a phase II detoxifying enzyme. It plays a role in the detoxification of environmental carcinogen metabolites, suggesting that its downregulation may contribute to HCC carcinogenesis secondary to ROS-facilitated oxidative damage. 51 The expression of GSTM1 was much higher in the KM and silymarin treatment groups versus control and DEN groups. Eclipta alba , a constitiuent of KM has been to shown to have antiproliferative property in HepG2 cell lines. Eclipta alba extract could induce DNA fragmentation and thus apoptosis in hepatic cell lines. 52 Alkaloids present in Eclipta alba can damage the DNA of cancer cells. 53 Eclipta alba is a well-known liver regenerative herb with protective effect on liver parenchymal cells and its cell membrane resulting in a decrease in enzyme seepage. 54 An important constituent of Terminalia chebula is chebulic acid. 55 Chebulic acid has been established to have hepatoprotective and antioxidant effects in various liver disease models. 56 In a previous study, Terminalia chebula stimulated apoptosis with membrane bleb and apoptotic bodies, thus rendering protection to macromolecules in DEN model by stabilizing the redox balance. 57 Piperine, present in Piper nigrum of Kadukkai maathirai has demonstrated dose-dependent cytotoxicity against HepG2 cells. It inhibits catalase and induces mitochondria-mediated cell death by H 2 O 2 in these cells. Further, analysis revealed that it had receptor tyrosine kinase inhibition property which led to inhibition of HCC progression. 58 Glucose, a major source of energy for cancer cells, is transported into cells via glucose transporters (GLUTs). These transporters are overexpressed in cancer cells which helps in enhanced glucose uptake by the cancer cells. Studies have shown that D-allose inhibited GLUT1 expression in cell lines of hepatocellular carcinoma thus reducing glucose utilization by cancer cells. 59 Citrus fruits contain valuable bioflavonoids that have bioactivities on apoptosis induction in vitro. 60 The phytochemicals present in citrus fruits exhibit antioxidant, anticancer, and anti-inflammatory activities. 61 – 63 The constituent plants of KM could have exerted synergistic effects to protect against DEN induced hepatic damage. Constitutive Wnt signalling has been document as a key factor in the development of various cancers. β catenin is a multifunctional protein in Wnt signalling pathway. Transcription factor 4 (Tcf4) is a peptide that interacts with the armadillo repeat region of β catenin leading to production of downstream transcription factors for malignant growth. The Tcf4 interaction regions on β catenin have been studied and used as templates for analysis. 64 , 65 Among the phytoconstituents of KM, D-allose contributed a maximum number of hydrogen interactions-4 with Lys435, His470. Octadecadienoicacid and propylene glycol monooleate showed the maximum number of interactions of eight with one hydrogen interaction. All other phytoconstituents showed interactions with this region of β catenin. With this analysis, we hypothesize that these constituents could interact with this region simultaneously or in part, thus potentially modulating the transcription process by preventing Tcf4 interaction with β catenin. We also noted that the shortlisted constituents would not modulate the LRP6-DKK1 mediated Wnt stimulation and subsequent pathway activation as they did not make any interactions with the LRP6-E3 binding site. In the present study beta sitosterol was confirmed in the GCMS analysis of KM. Earlier studies have reported that beta sitosterol downregulates beta catenin expression inhibits growth and induces apoptosis in colon cancer cells. 66 We presume that the active phytoconstituents of Kadukkai maathirai could modulate the intracellular transcription process by inhibiting the armadillo repeat region of β catenin. Conclusion We have concluded that Kadukkai maathirai could be potentially useful as a supplement in therapy of hepatocellular carcinoma. Further studies are needed to investigate its mechanism of action including validation of its target in hepatocellular carcinoma before exploring its efficacy in clinical studies. Ethics statement Seven-week-old female Sprague Dawley rats (150-200 g) procured from the Central Animal Research Facilities (CARF) of Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal were used in the study after getting approval from the Institutional Animal Ethics Committee approval (IAEC/KMC/19/2016 dated 16.03.2016). Guidelines given by the Committee for Control and Supervision of Experiments on Animals, Government of India, New Delhi for the use of laboratory animals were followed for the maintenance of animals. 18 Data availability Underlying data • Figshare: Macroscopic appearance of the liver https://doi.org/10.6084/m9.figshare.25117424.v1 67 • Figshare: Histopathology https://doi.org/10.6084/m9.figshare.25117436.v1 68 • Figshare: Polyacrylamide Gel Electrophoresis for the separation of proteins and mass spectrometry for the identification of proteins https://doi.org/10.6084/m9.figshare.25117442.v1 69 • Figshare: In-silico analysis of interactions of phytoconstituents of KM with β catenin https://doi.org/10.6084/m9.figshare.25117454.v1 70 • Figshare: Gas chromatography-mass spectroscopy (GC-MS) analysis of test drug KM https://doi.org/10.6084/m9.figshare.25117460.v1 71 • Figshare: Effect of KM on liver function in DEN-induced HCC model in rats https://doi.org/10.6084/m9.figshare.25117469.v1 72 • Figshare: Effect of “KM on body weight, relative weight of liver and nodule count in DEN-induced hepatocellular carcinoma model in rats https://doi.org/10.6084/m9.figshare.25117484.v1 73 • Figshare: Shortlisted phytoconstituents and their interactions with β catenin https://doi.org/10.6084/m9.figshare.25117490.v1 74 • Figshare: Gas chromatography-mass spectroscopy (GC-MS) analysis of the test drug Kadukkai maathirai (KM) https://doi.org/10.6084/m9.figshare.25117493.v1 75 • Figshare: Raw Data https://doi.org/10.6084/m9.figshare.25815934.v1 76 Reporting guidelines Figshare: ARRIVE checklist for Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study https://doi.org/10.6084/m9.figshare.25910563.v1 77 (Shetty MS. ARRIVE CHECKLIST [Internet]. figshare; 2024 [cited 2024 Jun 7]. Available from: https://figshare.com/articles/dataset/ARRIVE_CHECKLIST/25910563/1 ) Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0). Acknowledgments The authors would like to thank Manipal Academy of Higher Education, Manipal for providing the facilities to carry out the research. References 1. McGlynn KA, Petrick JL, London WT: Global Epidemiology of Hepatocellular Carcinoma: An Emphasis on Demographic and Regional Variability. Clin. Liver Dis. 2015; 19 : 223–238. PubMed Abstract | Publisher Full Text | Free Full Text 2. Singal AG, El-Serag HB: Hepatocellular Carcinoma From Epidemiology to Prevention: Translating Knowledge into Practice. Clin. Gastroenterol. Hepatol. 2015; 13 : 2140–2151. PubMed Abstract | Publisher Full Text | Free Full Text 3. Sachdeva M, Chawla YK, Arora SK: Immunology of hepatocellular carcinoma. World J. Hepatol. 2015; 7 : 2080–2090. PubMed Abstract | Publisher Full Text | Free Full Text 4. Grandhi MS, Kim AK, Ronnekleiv-Kelly SM, et al. : Hepatocellular carcinoma: From diagnosis to treatment. Surg. Oncol. 2016; 25 : 74–85. Publisher Full Text 5. op den Winkel M , Nagel D, Sappl J, et al. : Prognosis of Patients with Hepatocellular Carcinoma. Validation and Ranking of Established Staging-Systems in a Large Western HCC-Cohort. PLoS One. 2012; 7 : e45066. PubMed Abstract | Publisher Full Text | Free Full Text 6. Bellmunt J, Eisen T, Fishman M, et al. : Experience with sorafenib and adverse event management. Crit. Rev. Oncol. Hematol. 2011; 78 : 24–32. Publisher Full Text 7. Mazzoccoli G, Miele L, Oben J, et al. : Biology, Epidemiology, Clinical Aspects of Hepatocellular Carcinoma and the Role of Sorafenib. Curr. Drug Targets. 2016; 17 : 783–799. PubMed Abstract | Publisher Full Text 8. Fitzmorris P, Shoreibah M, Anand BS, et al. : Management of hepatocellular carcinoma. J. Cancer Res. Clin. Oncol. 2015; 141 (5): 861–876. Publisher Full Text 9. Therapeutic Index Siddha (Hospital Pharmacopoeia). SKM Siddha and ayurveda company; 2010. 10. Amuthan A: Physico-chemical evaluation of kadukkai maathirai and its tablet formulation, a Siddha iron preparation used in anemia.2012; 1 (1): 3–8. 11. Wagner H, Geyer B, Kiso Y, et al. : Coumestans as the main active principles of the liver drugs Eclipta alba and Wedelia calendulacea. Planta Med. 1986; 52 : 370–374. Publisher Full Text 12. Vijayakumar RS, Surya D, Nalini N: Antioxidant efficacy of black pepper (Piper nigrum L.) and piperine in rats with high fat diet induced oxidative stress. Redox Rep. 2004; 9 (2): 105–110. PubMed Abstract | Publisher Full Text 13. Wang YH, Morris-Natschke SL, Yang J, et al. : Anticancer Principles from Medicinal Piper (胡椒 Hú Jiāo) Plants. J. Tradit. Complement. Med. 2014; 4 (1): 8–16. PubMed Abstract | Publisher Full Text | Free Full Text 14. Selvendiran K, Sakthisekaran D: Chemopreventive effect of piperine on modulating lipid peroxidation and membrane bound enzymes in benzo(a) pyrene induced lung carcinogenesis. Biomed. Pharmacother. 2004; 58 (4): 264–267. PubMed Abstract | Publisher Full Text 15. Shetty M, Shenoy S, Kumar N, et al. : Kadukkai maathirai (A polyherbal traditional siddha formulation) prevents d-galactosamine induced hepatic necrosis in rats. Indian J. Tradit. Knowl. 2020; 19 (4): 708–713. 16. Verna L: N-Nitrosodiethylamine mechanistic data and risk assessment: Bioactivation, DNA-adduct formation, mutagenicity, and tumor initiation. Pharmacol. Ther. 1996; 71 (1-2): 57–81. PubMed Abstract | Publisher Full Text 17. Kerdput V, Nilbu-nga C, Kaewnoonual N, et al. : Therapeutic efficacy of a Dioscorea membranacea extract in a rat model of hepatocellular carcinoma: Histopathological aspects. J. Tradit. Complement. Med. 2021; 11 (5): 400–408. PubMed Abstract | Publisher Full Text | Free Full Text 18. Chief Editor: Government of india notifies the rules for breeding of and conducting animal experiments. Indian J. Pharm. 1999; 31 (1): 92–95. 19. Thomas S, Paul J, Shrungeswara A, et al. : Annona muricata fruit extract protects against diethylnitrosamine-induced hepatocellular cancer in rats. Asian Pac. J. Trop. Med. 2019; 12 (6): 272. Publisher Full Text 20. Jayakumar S, Madankumar A, Asokkumar S, et al. : Potential preventive effect of carvacrol against diethylnitrosamine-induced hepatocellular carcinoma in rats. Mol. Cell. Biochem. 2012; 360 (1-2): 51–60. PubMed Abstract | Publisher Full Text 21. Babiker R, Elmusharaf K, Keogh MB, et al. : Effect of Gum Arabic (Acacia Senegal) supplementation on visceral adiposity index (VAI) and blood pressure in patients with type 2 diabetes mellitus as indicators of cardiovascular disease (CVD): a randomized and placebo-controlled clinical trial. Lipids Health Dis. 2018; 17 (1): 56. PubMed Abstract | Publisher Full Text | Free Full Text 22. Kadukkai maathirai (a polyherbal traditional siddha formulation) prevents D-galactosamine induced hepatic necrosis in rats. Indian J. Tradit. Knowl. 2020; 19 (4). Publisher Full Text 23. Pradeep K, Victor Raj Mohan C, Gobianand K, et al. : Silymarin: An Effective Hepatoprotective Agent Against Diethylnitrosamine-Induced Hepatotoxicity in Rats. Pharm. Biol. 2007; 45 (9): 707–714. Publisher Full Text 24. Laferriere CA, Pang DS: Review of Intraperitoneal Injection of Sodium Pentobarbital as a Method of Euthanasia in Laboratory Rodents. J. Am. Assoc. Lab. Anim. Sci. 2020; 59 (3): 254–263. PubMed Abstract | Publisher Full Text | Free Full Text 25. Struck MB, Andrutis KA, Ramirez HE, et al. : Effect of a short-term fast on ketamine-xylazine anesthesia in rats. J. Am. Assoc. Lab. Anim. Sci. 2011; 50 (3): 344–348. PubMed Abstract 26. Thapa BR, Walia A: Liver function tests and their interpretation. Indian J. Pediatr. 2007; 74 (7): 663–671. Publisher Full Text 27. Titford M: The long history of hematoxylin. Biotech. Histochem. 2005; 80 (2): 73–78. PubMed Abstract | Publisher Full Text 28. Petrov A, Tsa A, Puglisi JD: Analysis of RNA by Analytical Polyacrylamide Gel Electrophoresis. Methods Enzymol. 2013; 530 : 301–313. PubMed Abstract | Publisher Full Text 29. Eberhardt J, Santos-Martins D, Tillack AF, et al. : AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings. J. Chem. Inf. Model. 2021; 61 (8): 3891–3898. PubMed Abstract | Publisher Full Text | Free Full Text 30. Trott O, Olson AJ: AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 2010; 31 (2): 455–461. PubMed Abstract | Publisher Full Text | Free Full Text 31. BIOVIA Discovery Studio Visualizer|Dassault Systèmes. Accessed April 4, 2024. Reference Source 32. Free Chemical Drawing Software for Students|ChemSketch|ACD/Labs. Accessed April 4, 2024. Reference Source 33. Padmakiran C, Prasad U, Koppala NS: Transdermal absorption studies of Nirgundi (Vitex Negundo Linn) taila and Kashaya (Decoction) with the aid of gas chromatography-mass spectrometry. J. Ayurveda. 2023; 17 (2): 79. Publisher Full Text 34. CLINICAL MANIFESTATIONS AND STUDIES IN PARENCHYMATOUS HEPATITIS. Ann. Intern. Med. 1937; 10 (8): 1144. Publisher Full Text 35. Barnes JM, Magee PN: Some Toxic Properties of Dimethylnitrosamine. Occup. Environ. Med. 1954; 11 (3): 167–174. PubMed Abstract | Publisher Full Text | Free Full Text 36. Magee PN, Barnes JM: The Production of Malignant Primary Hepatic Tumours in the Rat by Feeding Dimethylnitrosamine. Br. J. Cancer. 1956; 10 (1): 114–122. PubMed Abstract | Publisher Full Text | Free Full Text 37. Goldfarb S, Pugh TD, Koen H, et al. : Preneoplastic and neoplastic progression during hepatocarcinogenesis in mice injected with diethylnitrosamine in infancy. Environ. Health Perspect. 1983; 50 : 149–161. PubMed Abstract | Publisher Full Text | Free Full Text 38. Koen H, Pugh TD, Goldfarb S: Centrilobular distribution of diethylnitrosamine-induced hepatocellular foci in the mouse. Lab. Investig. 1983; 49 (1): 78–81. PubMed Abstract 39. Pascale RM, Simile MM, Feo F: Genomic abnormalities in hepatocarcinogenesis. Implications for a chemopreventive strategy. Anticancer Res. 13 (5A): 1341–1356. PubMed Abstract 40. Yang CS, Tu YY, Koop DR, et al. : Metabolism of nitrosamines by purified rabbit liver cytochrome P-450 isozymes. Cancer Res. 1985; 45 (3): 1140–1145. PubMed Abstract 41. Yoo JS, Guengerich FP, Yang CS: Metabolism of N-nitrosodialkylamines by human liver microsomes. Cancer Res. 1988; 48 (6): 1499–1504. PubMed Abstract 42. Thomas S, Paul J, Shrungeswara A, et al. : Annona muricata fruit extract protects against diethylnitrosamine-induced hepatocellular cancer in rats. Asian Pac. J. Trop. Med. 2019; 12 : 272. Publisher Full Text 43. Gupta C, Vikram A, Tripathi DN, et al. : Antioxidant and antimutagenic effect of quercetin against DEN induced hepatotoxicity in rat. Phytother. Res. 2010; 24 (1): 119–128. PubMed Abstract | Publisher Full Text 44. Singh D, Singh M, Yadav E, et al. : Attenuation of diethylnitrosamine (DEN) – Induced hepatic cancer in experimental model of Wistar rats by Carissa carandas embedded silver nanoparticles. Biomed. Pharmacother. 2018; 108 : 757–765. PubMed Abstract | Publisher Full Text 45. Bulle F, Mavier P, Zafrani ES, et al. : Mechanism of γ-glutamyl transpeptidase release in serum during intrahepatic and extrahepatic cholestasis in the rat: A histochemical, biochemical and molecular approach. Hepatology. 1990; 11 (4): 545–550. PubMed Abstract | Publisher Full Text 46. Lippert M, Papadopoulos N, Javadpour N: Role of lactate dehydrogenase isoenzymes in testicular cancer. Urology. 1981; 18 : 50–53. PubMed Abstract | Publisher Full Text 47. George J, Chandrakasan G: Biochemical abnormalities during the progression of hepatic fibrosis induced by dimethylnitrosamine. Clin. Biochem. 2000; 33 (7): 563–570. PubMed Abstract | Publisher Full Text 48. Abe R, Okano JI, Imamoto R, et al. : Sequential analysis of diethylnitrosamine-induced hepatocarcinogenesis in rats. Exp. Ther. Med. 2012; 3 (3): 371–378. PubMed Abstract | Publisher Full Text | Free Full Text 49. Panigrahy D, Edin ML, Lee CR, et al. : Epoxyeicosanoids stimulate multiorgan metastasis and tumor dormancy escape in mice. J. Clin. Invest. 2012; 122 : 178–191. PubMed Abstract | Publisher Full Text | Free Full Text 50. Xie C, Yi J, Lu J, et al. : N -Acetylcysteine Reduces ROS-Mediated Oxidative DNA Damage and PI3K/Akt Pathway Activation Induced by Helicobacter pylori Infection. Oxidative Med. Cell. Longev. 2018; 2018 : 1–9. PubMed Abstract | Publisher Full Text | Free Full Text 51. Lu Y, Zhou J, Zhang J, et al. : Dual roles of glutathione S-transferase mu 1 in the development and metastasis of hepatocellular carcinoma. Biomed. Pharmacother. 2019; 120 : 109532. Publisher Full Text 52. Chaudhary H, Dhuna V, Singh J, et al. : Evaluation of hydro-alcoholic extract of Eclipta alba for its anticancer potential: An in vitro study. J. Ethnopharmacol. 2011; 136 (2): 363–367. PubMed Abstract | Publisher Full Text 53. Abdel-Kader MS, Bahler BD, Malone S, et al. : DNA-damaging steroidal alkaloids from Eclipta alba from the suriname rainforest. J. Nat. Prod. 1998; 61 : 1202–1208. PubMed Abstract | Publisher Full Text 54. Chaudhary H, Jena PK, Seshadri S: In Vivo Evaluation of Eclipta alba Extract as Anticancer and Multidrug Resistance Reversal Agent. Nutr. Cancer. 2014; 66 (5): 904–913. Publisher Full Text 55. Juang LJ, Sheu SJ, Lin TC: Determination of hydrolyzable tannins in the fruit ofTerminalia chebula Retz. by high-performance liquid chromatography and capillary electrophoresis. J. Sep. Sci. 2004; 27 (9): 718–724. PubMed Abstract | Publisher Full Text 56. Lee HS, Jung SH, Yun BS, et al. : Isolation of chebulic acid from Terminalia chebula Retz. and its antioxidant effect in isolated rat hepatocytes. Arch. Toxicol. 2007; 81 : 211–218. PubMed Abstract | Publisher Full Text 57. Srigopalram S, Jayraaj IA: Effect of Terminalia chebula Retz on den induced hepatocellular carcinogenesis in experimental rats. Int. J. Pharm. Pharm. Sci. 2012. 58. Gunasekaran V, Elangovan K, Niranjali DS: Targeting hepatocellular carcinoma with piperine by radical-mediated mitochondrial pathway of apoptosis: An in vitro and in vivo study. Food Chem. Toxicol. 2017; 105 : 106–118. PubMed Abstract | Publisher Full Text 59. Noguchi C, Kamitori K, Hossain A, et al. : D-Allose Inhibits Cancer Cell Growth by Reducing GLUT1 Expression. Tohoku J. Exp. Med. 2016; 238 (2): 131–141. PubMed Abstract | Publisher Full Text 60. Banjerdpongchai R, Wudtiwai B, Khaw-on P, et al. : Hesperidin from Citrus seed induces human hepatocellular carcinoma HepG2 cell apoptosis via both mitochondrial and death receptor pathways. Tumour Biol. 2016; 37 (1): 227–237. PubMed Abstract | Publisher Full Text | Free Full Text 61. Chiba H, Uehara M, Wu J, et al. : Hesperidin, a Citrus Flavonoid, Inhibits Bone Loss and Decreases Serum and Hepatic Lipids in Ovariectomized Mice. J. Nutr. 2003; 133 (6): 1892–1897. PubMed Abstract | Publisher Full Text 62. Yumnam S, Park HS, Kim MK, et al. : Hesperidin Induces Paraptosis Like Cell Death in Hepatoblatoma, HepG2 Cells: Involvement of ERK1/2 MAPK. Calvisi D, ed. PLoS One. 2014; 9 (6): e101321. Publisher Full Text 63. Roohbakhsh A, Parhiz H, Soltani F, et al. : Molecular mechanisms behind the biological effects of hesperidin and hesperetin for the prevention of cancer and cardiovascular diseases. Life Sci. 2015; 124 : 64–74. PubMed Abstract | Publisher Full Text 64. Cheng Z, Biechele T, Wei Z, et al. : Crystal structures of the extracellular domain of LRP6 and its complex with DKK1. Nat. Struct. Mol. Biol. 2011; 18 (11): 1204–1210. PubMed Abstract | Publisher Full Text | Free Full Text 65. Poy F, Lepourcelet M, Shivdasani R, et al. : Structure of a human Tcf4–β-catenin complex. Nat. Struct. Mol. Biol. 2001; 8 : 1053–1057. PubMed Abstract | Publisher Full Text 66. Gu S, Liu F, Xie X, et al. : β-Sitosterol blocks the LEF-1-mediated Wnt/β-catenin pathway to inhibit proliferation of human colon cancer cells. Cell Signal. 2023 Apr; 104 : 110585. Epub 2023 Jan 2. PubMed Abstract | Publisher Full Text 67. Shetty MS: Macroscopic appearance of the liver.April 2024. Publisher Full Text 68. Shetty MS: Histopathology.April 2024. Publisher Full Text 69. Shetty MS: Polyacrylamide Gel Electrophoresis for the separation of proteins and mass spectrometry for the identification of proteins.April 2024. Publisher Full Text 70. Shetty MS: In-silico analysis of interactions of phytoconstituents of KM with β catenin.April 2024. Publisher Full Text 71. Shetty MS: Gas chromatography-mass spectroscopy (GC-MS) analysis of test drug KM.April 2024. Publisher Full Text 72. Shetty MS: Effect of KM on liver function in DEN-induced HCC model in rats.April 2024. Publisher Full Text 73. Shetty MS: Effect of “KM on body weight, relative weight of liver and nodule count in DEN-induced hepatocellular carcinoma model in rats”.April 2024. Publisher Full Text 74. Shetty MS: Shortlisted phytoconstituents and their interactions with β catenin.April 2024. Publisher Full Text 75. Shetty MS: Gas chromatography-mass spectroscopy (GC-MS) analysis of the test drug Kadukkai maathirai (KM).April 2024. Publisher Full Text 76. Shetty MS: Raw Data.2024. 77. Shetty MS: ARRIVE CHECKLIST.June 2024. Publisher Full Text Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 17 Jun 2024 ADD YOUR COMMENT Comment Author details Author details 1 Centre Of Excellence, Ocular Nanoscience, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 2 Division of Pharmacology, Department of Basic Medical Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 3 Department of Pharmacology, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 4 Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, Vaishali, Bihar, 844102, India 5 Department of Pharmacology, Manipal University College Malaysia, Bukit Baru, Melaka, 75150, Malaysia 6 Department of Cell and Molecular Biology, School of Life Sciences, Manipal Academy of Higher Education, Manipal, Manipal, Karnataka, 576104, India 7 Department of Research and Development, Muniyal Institute of Ayurveda and Medical Sciences, Manipal, Manipal, Karnataka, 576104, India 8 Division of Anatomy, Department of Basic Medical Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India Manjunath Shetty Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Smita Shenoy Roles: Project Administration, Supervision, Validation, Writing – Review & Editing Arul Amuthan Roles: Conceptualization, Methodology, Supervision Vasudha Devi Roles: Supervision Nitesh Kumar Roles: Methodology, Supervision, Validation, Visualization Amruth Kiran Roles: Resources, Software, Visualization, Writing – Review & Editing Ganesh Shenoy Roles: Investigation, Visualization, Writing – Review & Editing Diya Rajasekhar Chinta Roles: Software, Validation, Visualization, Writing – Review & Editing Shama Prasada K Roles: Software, Validation, Writing – Review & Editing Akshatha Shetty Roles: Validation, Visualization, Writing – Review & Editing Mohandas Rao K G Roles: Methodology, Resources, Validation, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (2) version 2 Revised Published: 22 Oct 2024, 13:639 https://doi.org/10.12688/f1000research.145961.2 version 1 Published: 17 Jun 2024, 13:639 https://doi.org/10.12688/f1000research.145961.1 Copyright © 2024 Shetty M et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Shetty M, Shenoy S, Amuthan A et al. Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.12688/f1000research.145961.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 2 VERSION 2 PUBLISHED 22 Oct 2024 Revised Views 0 Cite How to cite this report: Salama AF. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.172786.r345193 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v2#referee-response-345193 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 09 Jan 2025 Afrah Fatthi Salama , Professor of Biochemistry, Faculty of Science, Tanta University, Tanta, Gharbia Governorate, Egypt Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.172786.r345193 Page 5, under Animals, please add Reference for using female rats, giving reason for using female not male You gave DEN, in drinking water as Reference no. 20, this ... Continue reading READ ALL Page 5, under Animals, please add Reference for using female rats, giving reason for using female not male You gave DEN, in drinking water as Reference no. 20, this reference used male rats? Is that effect applied on female? And do you calculate the dose that every female rat received for tumor induction? Page 7, Table 2. Relative liver weight, how calculated?, you put (g)?, it have to be (g/100 g Body weight), okay. Also Body weight (g), what does it mean? Is that the final body weight?. It have to be (Body weight gain, which is the difference between animal weight at the beginning and at the end of the experiment , to be positive or negative number), please correct it as I explained. The same table 2. Nodule count, how can you do this count?. I noticed that the dose 36 mg/kg of KM is the best, than larger ones, also silymarin give the lowest number of nodule count than all doses of KM, and you have to explain these results well in the paper In fig. 1. why you did not mark the position of nodules like the other groups? Write Glutathione S- Transferase , like this Finally Results , Discussion and conclusion need more explanation for all results. Best Regards Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? I cannot comment. A qualified statistician is required. Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests: No competing interests were disclosed. Reviewer Expertise: Clinical Biochemistry- Natural products –Cancer Biology-Toxicology, and my Search inter in applied fields, see it on google scholar I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Salama AF. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.172786.r345193 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v2#referee-response-345193 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Version 1 VERSION 1 PUBLISHED 17 Jun 2024 Views 0 Cite How to cite this report: Emran TB. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309884 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309884 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 21 Aug 2024 Talha Bin Emran , BGC Trust University Bangladesh, Chittagong, Chittagong Division, Bangladesh Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.159988.r309884 1. The results and discussion section could benefit from a more detailed exploration of the findings. It would be helpful to provide additional insight into why certain effects are coming in to existence and what could be the possible reason behind ... Continue reading READ ALL 1. The results and discussion section could benefit from a more detailed exploration of the findings. It would be helpful to provide additional insight into why certain effects are coming in to existence and what could be the possible reason behind them? 2. Results and Conclusion: The section devoted to the explanation of the results suffers from the same problems revealed so far. The storyline in the results section (and conclusion) is hard to follow. Moreover, the conclusions presented appear to diverge from the insights suggested by the empirical results. It may be beneficial to revisit and align the conclusions more closely with the findings. 3. The discussion should be rather organized around arguments avoiding simply describing details without providing much meaning. A real discussion should also link the findings of the study to theory and/or literature. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Postdoctoral scientist with 2+ years of experience in viral and non-viral gene editing, particularly using the CRISPR/Cas9 system and DNA Damage Repair (DDR) in cancer biology. Proficient in handling mammalian cancer cells, conducting cell viability assays, and performing advanced molecular techniques such as next-generation sequencing. Actively engaged in researching DDR gene targets to overcome cancer therapy resistance. Prepared to contribute to the academic, pharmaceutical, biotech, and translational research sectors, focusing on gene therapy, gene editing, immune-oncology, mRNA, oncological biomarkers, and the development of small molecule/DDR-targeted inhibitors. Possesses a strong foundation in the practical applications and potential of small molecule inhibitor screening. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Emran TB. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309884 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309884 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Yu H. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309885 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309885 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 21 Aug 2024 Haiyang Yu , Tianjin University of Traditional Chinese Medicine, Tianjin, China Approved VIEWS 0 https://doi.org/10.5256/f1000research.159988.r309885 This article demonstrates that KM can protect the liver from further damage by reducing the nodule count and reducing the relative weight of the liver. Also, this paper verifies that KM regulates the intracellular transcription process by ... Continue reading READ ALL This article demonstrates that KM can protect the liver from further damage by reducing the nodule count and reducing the relative weight of the liver. Also, this paper verifies that KM regulates the intracellular transcription process by reducing GSTM1 expression and simultaneously inhibiting the armadillo repeat region of β catenin to realize the potential of inhibiting HCC. But the hepatoprotection mechanism of KM against diethylnitrosamine-induced liver cancer needs to be further improved. It suggest that the author should further study and explain. The arrangement of the pictures, tables, and text in the article is too messy. For example, Figure 5 and Table 4, it is recommended to arrange them separately. We recommended that the picture and tables should be placed above or below the page for reading. In some similar paper, some cell experiments will be done to verify the results obtained in animal experiments. We suggest that the author can do some cell experiments to further study. Page 16, conclusions, “Hence, it could be concluded that the collective effect of all the herbs present in KM was responsible for its beneficial effect.” The description is just too absolute. Please unify the cell line names included in the article. For example, page 15，HepG2 cell and Hep G2 cells. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: hepatocellular cancer progression I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Yu H. Reviewer Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309885 ) The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309885 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 17 Jun 2024 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 Version 2 (revision) 22 Oct 24 read Version 1 17 Jun 24 read read Haiyang Yu , Tianjin University of Traditional Chinese Medicine, Tianjin, China Talha Bin Emran , BGC Trust University Bangladesh, Chittagong, Bangladesh Afrah Fatthi Salama , Tanta University, Tanta, Egypt Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Salama A. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 09 Jan 2025 | for Version 2 Afrah Fatthi Salama , Professor of Biochemistry, Faculty of Science, Tanta University, Tanta, Gharbia Governorate, Egypt 0 Views copyright © 2025 Salama A. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Page 5, under Animals, please add Reference for using female rats, giving reason for using female not male You gave DEN, in drinking water as Reference no. 20, this reference used male rats? Is that effect applied on female? And do you calculate the dose that every female rat received for tumor induction? Page 7, Table 2. Relative liver weight, how calculated?, you put (g)?, it have to be (g/100 g Body weight), okay. Also Body weight (g), what does it mean? Is that the final body weight?. It have to be (Body weight gain, which is the difference between animal weight at the beginning and at the end of the experiment , to be positive or negative number), please correct it as I explained. The same table 2. Nodule count, how can you do this count?. I noticed that the dose 36 mg/kg of KM is the best, than larger ones, also silymarin give the lowest number of nodule count than all doses of KM, and you have to explain these results well in the paper In fig. 1. why you did not mark the position of nodules like the other groups? Write Glutathione S- Transferase , like this Finally Results , Discussion and conclusion need more explanation for all results. Best Regards Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? I cannot comment. A qualified statistician is required. Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Clinical Biochemistry- Natural products –Cancer Biology-Toxicology, and my Search inter in applied fields, see it on google scholar I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Salama AF. Peer Review Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.172786.r345193) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-639/v2#referee-response-345193 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Emran T. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 21 Aug 2024 | for Version 1 Talha Bin Emran , BGC Trust University Bangladesh, Chittagong, Chittagong Division, Bangladesh 0 Views copyright © 2024 Emran T. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions 1. The results and discussion section could benefit from a more detailed exploration of the findings. It would be helpful to provide additional insight into why certain effects are coming in to existence and what could be the possible reason behind them? 2. Results and Conclusion: The section devoted to the explanation of the results suffers from the same problems revealed so far. The storyline in the results section (and conclusion) is hard to follow. Moreover, the conclusions presented appear to diverge from the insights suggested by the empirical results. It may be beneficial to revisit and align the conclusions more closely with the findings. 3. The discussion should be rather organized around arguments avoiding simply describing details without providing much meaning. A real discussion should also link the findings of the study to theory and/or literature. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Postdoctoral scientist with 2+ years of experience in viral and non-viral gene editing, particularly using the CRISPR/Cas9 system and DNA Damage Repair (DDR) in cancer biology. Proficient in handling mammalian cancer cells, conducting cell viability assays, and performing advanced molecular techniques such as next-generation sequencing. Actively engaged in researching DDR gene targets to overcome cancer therapy resistance. Prepared to contribute to the academic, pharmaceutical, biotech, and translational research sectors, focusing on gene therapy, gene editing, immune-oncology, mRNA, oncological biomarkers, and the development of small molecule/DDR-targeted inhibitors. Possesses a strong foundation in the practical applications and potential of small molecule inhibitor screening. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Emran TB. Peer Review Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309884) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309884 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Yu H. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 21 Aug 2024 | for Version 1 Haiyang Yu , Tianjin University of Traditional Chinese Medicine, Tianjin, China 0 Views copyright © 2024 Yu H. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions This article demonstrates that KM can protect the liver from further damage by reducing the nodule count and reducing the relative weight of the liver. Also, this paper verifies that KM regulates the intracellular transcription process by reducing GSTM1 expression and simultaneously inhibiting the armadillo repeat region of β catenin to realize the potential of inhibiting HCC. But the hepatoprotection mechanism of KM against diethylnitrosamine-induced liver cancer needs to be further improved. It suggest that the author should further study and explain. The arrangement of the pictures, tables, and text in the article is too messy. For example, Figure 5 and Table 4, it is recommended to arrange them separately. We recommended that the picture and tables should be placed above or below the page for reading. In some similar paper, some cell experiments will be done to verify the results obtained in animal experiments. We suggest that the author can do some cell experiments to further study. Page 16, conclusions, “Hence, it could be concluded that the collective effect of all the herbs present in KM was responsible for its beneficial effect.” The description is just too absolute. Please unify the cell line names included in the article. For example, page 15，HepG2 cell and Hep G2 cells. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise hepatocellular cancer progression I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Yu H. Peer Review Report For: Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study [version 2; peer review: 1 approved, 2 approved with reservations] . F1000Research 2024, 13 :639 ( https://doi.org/10.5256/f1000research.159988.r309885) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-639/v1#referee-response-309885 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions Adjust parameters to alter display View on desktop for interactive features Includes Interactive Elements View on desktop for interactive features Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Stay Updated Sign up for content alerts and receive a weekly or monthly email with all newly published articles Register with F1000Research Already registered? Sign in Not now, thanks close PLEASE NOTE If you are an AUTHOR of this article, please check that you signed in with the account associated with this article otherwise we cannot automatically identify your role as an author and your comment will be labelled as a “User Comment”. If you are a REVIEWER of this article, please check that you have signed in with the account associated with this article and then go to your account to submit your report, please do not post your review here. If you do not have access to your original account, please contact us . All commenters must hold a formal affiliation as per our Policies . The information that you give us will be displayed next to your comment. User comments must be in English, comprehensible and relevant to the article under discussion. We reserve the right to remove any comments that we consider to be inappropriate, offensive or otherwise in breach of the User Comment Terms and Conditions . Commenters must not use a comment for personal attacks. When criticisms of the article are based on unpublished data, the data should be made available. I accept the User Comment Terms and Conditions Please confirm that you accept the User Comment Terms and Conditions. Affiliation ✕ refresh Please enter your institution. Note: To add your institution or organisation, start typing the name and then select the correct name from the list. Where applicable, the name will appear in both the original language and in English. Do not paste in the name. If the name does not appear in the drop-down list, we will display the information you have entered. ✕ refresh Country/Region * USA UK Canada China France Germany Afghanistan Aland Islands Albania Algeria American Samoa Andorra Angola Anguilla Antarctica Antigua and Barbuda Argentina Armenia Aruba Australia Austria Azerbaijan Bahamas Bahrain Bangladesh Barbados Belarus Belgium Belize Benin Bermuda Bhutan Bolivia Bosnia and Herzegovina Botswana Bouvet Island Brazil British Indian Ocean Territory British Virgin Islands Brunei Bulgaria Burkina Faso Burundi Cambodia Cameroon Canada Cape Verde Cayman Islands Central African Republic Chad Chile China Christmas Island Cocos (Keeling) Islands Colombia Comoros Congo Cook Islands Costa Rica Cote d'Ivoire Croatia Cuba Cyprus Czech Republic Democratic Republic of the Congo Denmark Djibouti Dominica Dominican Republic Ecuador Egypt El Salvador Equatorial Guinea Eritrea Estonia Ethiopia Falkland Islands Faroe Islands Federated States of Micronesia Fiji Finland France French Guiana French Polynesia French Southern Territories Gabon Georgia Germany Ghana Gibraltar Greece Greenland Grenada Guadeloupe Guam Guatemala Guernsey Guinea Guinea-Bissau Guyana Haiti Heard Island and Mcdonald Islands Holy See (Vatican City State) Honduras Hong Kong Hungary Iceland India Indonesia Iran Iraq Ireland Israel Italy Jamaica Japan Jersey Jordan Kazakhstan Kenya Kiribati Kosovo (Serbia and Montenegro) Kuwait Kyrgyzstan Lao People's Democratic Republic Latvia Lebanon Lesotho Liberia Libya Liechtenstein Lithuania Luxembourg Macao Madagascar Malawi Malaysia Maldives Mali Malta Marshall Islands Martinique Mauritania Mauritius Mayotte Mexico Minor Outlying Islands of the United States Moldova Monaco Mongolia Montenegro Montserrat Morocco Mozambique Myanmar Namibia Nauru Nepal Netherlands Antilles New Caledonia New Zealand Nicaragua Niger Nigeria Niue Norfolk Island North Korea North Macedonia Northern Mariana Islands Norway Oman Pakistan Palau Palestinian Territory Panama Papua New Guinea Paraguay Peru Philippines Pitcairn Poland Portugal Puerto Rico Qatar Reunion Romania Russian Federation Rwanda Saint Helena Saint Kitts and Nevis Saint Lucia Saint Pierre and Miquelon Saint Vincent and the Grenadines Samoa San Marino Sao Tome and Principe Saudi Arabia Senegal Serbia Seychelles Sierra Leone Singapore Slovakia Slovenia Solomon Islands Somalia South Africa South Georgia and the South Sandwich Is South Korea South Sudan Spain Sri Lanka Sudan Suriname Svalbard and Jan Mayen Swaziland Sweden Switzerland Syria Taiwan Tajikistan Tanzania Thailand The Gambia The Netherlands Timor-Leste Togo Tokelau Tonga Trinidad and Tobago Tunisia Turkey Turkmenistan Turks and Caicos Islands Tuvalu UK USA Uganda Ukraine United Arab Emirates United States Virgin Islands Uruguay Uzbekistan Vanuatu Venezuela Vietnam Wallis and Futuna West Bank and Gaza Strip Western Sahara Yemen Zambia Zimbabwe Please select your country/region. You must enter a comment. Competing Interests Please disclose any competing interests that might be construed to influence your judgment of the article's or peer review report's validity or importance. Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Please state your competing interests The comment has been saved. An error has occurred. Please try again. Cancel Post var lTitle = "Kadukkai maathirai (Indian herbal drug) prevents...".replace("'", ''); var linkedInUrl = "http://www.linkedin.com/shareArticle?url=https://f1000research.com/articles/13-639/v2" + "&title=" + encodeURIComponent(lTitle) + "&summary=" + encodeURIComponent('Read the article by '); var deliciousUrl = "https://del.icio.us/post?url=https://f1000research.com/articles/13-639/v2&title=" + encodeURIComponent(lTitle); var redditUrl = "http://reddit.com/submit?url=https://f1000research.com/articles/13-639/v2" + "&title=" + encodeURIComponent(lTitle); linkedInUrl += encodeURIComponent('Shetty M et al.'); var offsetTop = /chrome/i.test( navigator.userAgent ) ? 4 : -10; var addthis_config = { ui_offset_top: offsetTop, services_compact : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_expanded : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_custom : [ { name: "LinkedIn", url: linkedInUrl, icon:"/img/icon/at_linkedin.svg" }, { name: "Mendeley", url: "http://www.mendeley.com/import/?url=https://f1000research.com/articles/13-639/v2/mendeley", icon:"/img/icon/at_mendeley.svg" }, { name: "Reddit", url: redditUrl, icon:"/img/icon/at_reddit.svg" }, ] }; var addthis_share = { url: "https://f1000research.com/articles/13-639", templates : { twitter : "Kadukkai maathirai (Indian herbal drug) prevents hepatocellular.... Shetty M et al., published by " + "@F1000Research" + ", https://f1000research.com/articles/13-639/v2" } }; if (typeof(addthis) != "undefined"){ addthis.addEventListener('addthis.ready', checkCount); addthis.addEventListener('addthis.menu.share', checkCount); } $(".f1r-shares-twitter").attr("href", "https://twitter.com/intent/tweet?text=" + addthis_share.templates.twitter); $(".f1r-shares-facebook").attr("href", "https://www.facebook.com/sharer/sharer.php?u=" + addthis_share.url); $(".f1r-shares-linkedin").attr("href", addthis_config.services_custom[0].url); $(".f1r-shares-reddit").attr("href", addthis_config.services_custom[2].url); $(".f1r-shares-mendelay").attr("href", addthis_config.services_custom[1].url); function checkCount(){ setTimeout(function(){ $(".addthis_button_expanded").each(function(){ var count = $(this).text(); if (count !== "" && count != "0") $(this).removeClass("is-hidden"); else $(this).addClass("is-hidden"); }); }, 1000); } close How to cite this report {{reportCitation}} Cancel Copy Citation Details $(function(){R.ui.buttonDropdowns('.dropdown-for-downloads');}); $(function(){R.ui.toolbarDropdowns('.toolbar-dropdown-for-downloads');}); $.get("/articles/acj/145961/172786") new F1000.Clipboard(); new F1000.ThesaurusTermsDisplay("articles", "article", "172786"); $(document).ready(function() { $( "#frame1" ).on('load', function() { var mydiv = $(this).contents().find("div"); var h = mydiv.height(); console.log(h) }); var tooltipLivingFigure = jQuery(".interactive-living-figure-label .icon-more-info"), titleLivingFigure = tooltipLivingFigure.attr("title"); tooltipLivingFigure.simpletip({ fixed: true, position: ["-115", "30"], baseClass: 'small-tooltip', content:titleLivingFigure + " " }); tooltipLivingFigure.removeAttr("title"); $("body").on("click", ".cite-living-figure", function(e) { e.preventDefault(); var ref = $(this).attr("data-ref"); $(this).closest(".living-figure-list-container").find("#" + ref).fadeIn(200); }); $("body").on("click", ".close-cite-living-figure", function(e) { e.preventDefault(); $(this).closest(".popup-window-wrapper").fadeOut(200); }); $(document).on("mouseup", function(e) { var metricsContainer = $(".article-metrics-popover-wrapper"); if (!metricsContainer.is(e.target) && metricsContainer.has(e.target).length === 0) { $(".article-metrics-close-button").click(); } }); var articleId = $('#articleId').val(); if($("#main-article-count-box").attachArticleMetrics) { $("#main-article-count-box").attachArticleMetrics(articleId, { articleMetricsView: true }); } }); var figshareWidget = $(".new_figshare_widget"); if (figshareWidget.length > 0) { window.figshare.load("f1000", function(Widget) { // Select a tag/tags defined in your page. In this tag we will place the widget. _.map(figshareWidget, function(el){ var widget = new Widget({ articleId: $(el).attr("figshare_articleId") //height:300 // this is the height of the viewer part. [Default: 550] }); widget.initialize(); // initialize the widget widget.mount(el); // mount it in a tag that's on your page // this will save the widget on the global scope for later use from // your JS scripts. This line is optional. //window.widget = widget; }); }); } close Error Close Add Reset F1000.MICROSERVICES.AFFILIATION = ''; $(document).ready(function () { $('.js-affiliations-form').each((index, form) => { new AffiliationForm({ formId: form.id, institutionErrorSelector: '.comment-enter-institution', departmentErrorSelector: '.comment-enter-department', placeSelector: '.js-add-comment-place', stateSelector: '.js-add-comment-state', zipCodeSelector: '.js-add-comment-zipcode', countrySelector: '.js-add-comment-country', countryErrorSelector: '.comment-enter-country', }); }); }); $(document).ready(function () { var reportIds = { "338561": 0, "338560": 0, "333853": 0, "333852": 0, "334797": 0, "334796": 0, "334799": 0, "334798": 0, "334805": 0, "334804": 0, "294741": 0, "342103": 0, "294742": 0, "342102": 0, "294743": 0, "334801": 0, "334800": 0, "334803": 0, "334802": 0, "342109": 0, "294748": 0, "342108": 0, "294749": 0, "342111": 0, "294750": 0, "342110": 0, "342105": 0, "294744": 0, "342104": 0, "294745": 0, "342107": 0, "294746": 0, "342106": 0, "294747": 0, "345191": 0, "345190": 0, "345197": 0, "345196": 0, "345199": 0, "345198": 0, "345193": 6, "345192": 0, "345195": 0, "345194": 0, "309878": 0, "309879": 0, "338557": 0, "309884": 26, "338556": 0, "309885": 24, "338559": 0, "309886": 0, "338558": 0, "309887": 0, "338553": 0, "309880": 0, "338552": 0, "309881": 0, "338555": 0, "309882": 0, "338554": 0, "309883": 0, }; $(".referee-response-container,.js-referee-report").each(function(index, el) { var reportId = $(el).attr("data-reportid"), reportCount = reportIds[reportId] || 0; $(el).find(".comments-count-container,.js-referee-report-views").html(reportCount); }); var uuidInput = $("#article_uuid"), oldUUId = uuidInput.val(), newUUId = "6e7dc4c0-641c-4dc0-9c50-b8ecaa6891c3"; uuidInput.val(newUUId); $("a[href*='article_uuid=']").each(function(index, el) { var newHref = $(el).attr("href").replace(oldUUId, newUUId); $(el).attr("href", newHref); }); }); An innovative open access publishing platform offering rapid publication and open peer review, whilst supporting data deposition and sharing. Browse Gateways Collections How it Works Contact For Developers Cookie Notice Privacy Notice RSS Submit Your Research Follow us © 2012-2026 F1000 Research Ltd. ISSN 2046-1402 | Legal | Partner of Research4Life • CrossRef • ORCID • FAIRSharing R.templateTests.simpleTemplate = R.template(' $text $text $text $text $text '); R.templateTests.runTests(); var F1000platform = new F1000.Platform({ name: "f1000research", displayName: "F1000Research", hostName: "f1000research.com", id: "1", editorialEmail: "[email protected]", infoEmail: "[email protected]", usePmcStats: true }); $(function(){R.ui.dropdowns('.dropdown-for-authors, .dropdown-for-about, .dropdown-for-myresearch');}); // $(function(){R.ui.dropdowns('.dropdown-for-referees');}); $(document).ready(function () { if ($(".cookie-warning").is(":visible")) { $(".sticky").css("margin-bottom", "35px"); $(".devices").addClass("devices-and-cookie-warning"); } $(".cookie-warning .close-button").click(function (e) { $(".devices").removeClass("devices-and-cookie-warning"); $(".sticky").css("margin-bottom", "0"); }); $("#tweeter-feed .tweet-message").each(function (i, message) { var self = $(message); self.html(linkify(self.html())); }); $(".partner").on("mouseenter mouseleave", function() { $(this).find(".gray-scale, .colour").toggleClass("is-hidden"); }); }); Sign In Remember me Forgotten your password? Sign In Cancel Email or password not correct. Please try again Please wait... $(function(){ // Note: All the setup needs to run against a name attribute and *not* the id due the clonish // nature of facebox... $("a[id=googleSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("GOOGLE"); $("form[id=oAuthForm]").submit(); }); $("a[id=facebookSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("FACEBOOK"); $("form[id=oAuthForm]").submit(); }); $("a[id=orcidSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("ORCID"); $("form[id=oAuthForm]").submit(); }); }); If you've forgotten your password, please enter your email address below and we'll send you instructions on how to reset your password. The email address should be the one you originally registered with F1000. Email address not valid, please try again You registered with F1000 via Google, so we cannot reset your password. To sign in, please click here . If you still need help with your Google account password, please click here . You registered with F1000 via Facebook, so we cannot reset your password. To sign in, please click here . If you still need help with your Facebook account password, please click here . Code not correct, please try again Reset password Cancel Email us for further assistance. Server error, please try again. If your email address is registered with us, we will email you instructions to reset your password. If you think you should have received this email but it has not arrived, please check your spam filters and/or contact for further assistance. Please wait... Register $(document).ready(function () { signIn.createSignInAsRow($("#sign-in-form-gfb-popup")); $(".target-field").each(function () { var uris = $(this).val().split("/"); if (uris.pop() === "login") { $(this).val(uris.toString().replace(",","/")); } }); });