Molecular biomarkers in type 2 diabetes and colorectal cancer

Globally, Type 2 Diabetes (T2D) and Colorectal Cancer (CRC) are among the most prevalent metabolic diseases and cancers, respectively. T2D is a progressive disease encompassing two underlying progressive conditions, insulin resistance and pancreas functional β-cell loss. CRC mostly arise from the aberrant proliferation of colonic epithelial cells in the form of colorectal polyps. T2D and CRC are chronic diseases and understanding the underlying molecular mechanism to identify biomarkers of their progression could help prevent or delay the course of the two diseases. Additionally, obesity is the major risk factor of T2D, and they are both considered as CRC risk factors. Therefore, there are common factors involved in the progression of T2D and CRC. In this regard, my thesis aimed to investigate T2D and CRC progression in four sections. Telomere length shortening is one of the hallmarks of cellular senescence. Shorter leukocyte telomere length in T2D has been reported previously. Firstly, I aimed to investigate cellular senescence and assess telomere biology using a cell model of human pancreas islet-derived progenitor cells (hIPCs). The second section studied was on how T2D progression was related to gut hormones regulating the insulin secretion from pancreas islets. Thirdly, SCFA was studied in relation to CRC progression. By conducting a systematic review and meta-analysis, a link between faecal SCFA level and the risk and incidence of CRC was established. Finally, in the fourth section, the concerning rising incidence of early-onset CRC (EOCRC: CRC diagnosed earlier than 50 years of age) led us to investigate this further by analysing a consecutive series of 3609 patients that underwent colorectal surgery over a 26-year period at Concord Hospital (NSW, Australia). The focus was on the incidence trend, clinicopathological features and survival in this age group. The current National Bowel Cancer Screening Program commences at the age of 50 years. Our findings including a significantly higher percentage of metastasis in EOCRC, as well as significantly higher risk of developing an advanced tumour in this age group provides supporting evidence for lowering the commencement age for CRC screening

Short-chain fatty acid concentrations in the incidence and risk-stratification of colorectal cancer:a systematic review and meta-analysis

BACKGROUND: The beneficial role of gut microbiota and bacterial metabolites, including short-chain fatty acids (SCFAs), is well recognized, although the available literature around their role in colorectal cancer (CRC) has been inconsistent. METHODS: We performed a systematic review and meta-analysis to examine the associations of fecal SCFA concentrations to the incidence and risk of CRC. Data extraction through Medline, Embase, and Web of Science was carried out from database conception to June 29, 2022. Predefined inclusion/exclusion criteria led to the selection of 17 case-control and six cross-sectional studies for quality assessment and analyses. Studies were categorized for CRC risk or incidence, and RevMan 5.4 was used to perform the meta-analyses. Standardized mean differences (SMD) with 95% confidence intervals (CI) were calculated using a random-effects model. Studies lacking quantitation were included in qualitative analyses. RESULTS: Combined analysis of acetic, propionic, and butyric acid revealed significantly lower concentrations of these SCFAs in individuals with a high-risk of CRC (SMD = 2.02, 95% CI 0.31 to 3.74, P = 0.02). Additionally, CRC incidence was higher in individuals with lower levels of SCFAs (SMD = 0.45, 95% CI 0.19 to 0.72, P = 0.0009), compared to healthy individuals. Qualitative analyses identified 70.4% of studies reporting significantly lower concentrations of fecal acetic, propionic, butyric acid, or total SCFAs in those at higher risk of CRC, while 66.7% reported significantly lower concentrations of fecal acetic and butyric acid in CRC patients compared to healthy controls. CONCLUSIONS: Overall, lower fecal concentrations of the three major SCFAs are associated with higher risk of CRC and incidence of CRC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12916-022-02529-4

Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori

UreB is one of the urease subunits of Helicobacter pylori and can be used as an excellent antigen candidate for H. pylori vaccination. Easy access to highly purified UreB protein, facilitate advances in therapeutic or preventive strategies. To achieve a simplified purification procedure, the present report represents a novel method of producing recombinant urease subunit B extracellularly. ureB gene from 26,695 standard strain was amplified by PCR and cloned into pET-26b(+) expression vector. UreB was expressed as a soluble, N-terminal pelB and C-terminal hexahistidine-tagged fusion protein (UreB-6His) and secreted into the periplasmic space of Escherichia coli. Expression of the recombinant UreB in E. coli BL21 (DE3) was induced by isopropylthio-β-d-galactoside (IPTG). Expression of UreB was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blot analysis using anti-His monoclonal antibody. UreB-6His protein was extracted from the periplasm by osmotic shock treatment and was purified in one step by Nickel affinity chromatography. In conclusion, the present protocol is easier to perform; more time effective and low cost than earlier methods. Keywords Helicobacter pylori –Urease subunit B–Cloning–Periplasmic expression Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori (PDF Download Available). Available from: https://www.researchgate.net/publication/227181089_Periplasmic_expression_and_one-step_purification_of_urease_subunit_B_of_Helicobacter_pylori [accessed Dec 09 2017]

Expression and Purification of the Recombinant Cytochrome P450 CYP141 Protein of Mycobacterium Tuberculosis as a Diagnostic Tool and Vaccine Production

Background: Tuberculosis (TB) is regarded as a health problem worldwide, particularly in developing countries. Mycobacterium tuberculosis (M. tuberculosis) is the cause of this disease. Approximately two billion people worldwide are infected by M. tuberculosis and annually about two million individuals die in consequence. Forty million people are estimated to die because of M. tuberculosis over the next 25 years if the measures for controlling this infection are not extensively developed. In the vaccination field, Bacillus Calmette-Guerin (BCG) is still the most effective vaccine but it shows no efficacy in adult pulmonary patients. One of the other problems regarding TB is its appropriate diagnosis. Objectives: In this experimental study, the recombinant cytochrome P450 CYP141 protein of M. tuberculosis was expressed and purified to be used as a vaccine candidate and diagnostic purpose in subsequent investigations. Materials and Methods: The optimization of the cytochrome P450 CYP141 protein expression was evaluated in different conditions. Then, this protein was purified with a resin column of nickel-nitrilotriacetic acid and investigated via Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western Blotting. Results: The highest expression of the cytochrome P450 CYP141 protein was obtained by the addition of 1 mM of isopropyl beta-D-1-thiogalactopyranoside (IPTG) to the bacterial culture grown to an optical density at 600 nm (OD600) of 0.6, 16 hours after induction. This protein was subsequently purified with a purification of higher than 80%. The results of Western Blotting indicated that the purified protein was specifically detected. Conclusions: In this experimental study, for the first time in Iran the expression and purification of this recombinant protein was done successfully. This recombinant protein could be used as a vaccine candidate and diagnostic purpose in subsequent investigations

Vitamin D suppresses cellular pathways of diabetes complication in liver

Objective(s): The aim of this study was to investigate the effect of vitamin D on glucose metabolism, as well as the expression of five key genes involved in the development of diabetes complications in liver tissue of diabetic rats. Materials and Methods: Twenty-four male Sprague–Dawley rats were randomly divided into three groups (8 rats in each group). The first group served as control and the other two groups received an intraperitoneal injection of 45 mg/kg streptozotocin to develop diabetes. Groups were treated for four weeks either with placebo or vitamin D (two injections of 20000 IU/kg). Thereafter, serum levels of glucose, insulin and HbA1c were assessed. Liver tissue was examined for the level of advanced glycation end products (AGEs) and the gene expression of AGE cellular receptor (AGER), glyoxalase-1 (GLO-1), aldose reductase (AR), O-linked N-acetylglucosamine transferase (OGT) and glutamine/ fructose-6-phosphate aminotransferase (GFAT). Results: Vitamin D injection resulted in a significant increase in plasma level of 25-hydroxycholecalciferol, which could improve hyperglycemia about 11% compared to placebo-receiving diabetic rats (P=0.005). Insulin level increased as a result of vitamin D treatment compared to control (3.31±0.65 vs. 2.15±0.79; P= 0.01). Serum HbA1c and liver AGE concentrations had a slight but insignificant reduction following vitamin D intake. Moreover, a significant decline was observed in gene expression of AGER and OGT in liver tissue (P=0.04 and PConclusion: Vitamin D might contribute in ameliorating diabetes complications not only by improving blood glucose and insulin levels, but also by suppressing AGER and OGT gene expression in the liver

LED visible light assisted photo-oxidation of acetaminophen using one-step synthesis of Cu,Fe@g-C3N4 nanosheet – Activated persulfate system in aqueous solutions

In this work, the synthesis of Iron (Fe) and Copper (Cu) co-doped g-C3N4 was performed using the thermal decomposition of urea while iron nitrate and copper nitrate were used as dopant precursors. The fabricated catalyst (Fe-Cu@g-C3N4) was coupled with visible light and used for acetaminophen (APAP) degradation. The synthesized catalyst was characterized via several techniques including XRD, BET, BJH, SEM, TEM, EDX, EDS Dot mapping, DLS, and UV–Vis deflective reflectance spectroscopy. The performed characterization tests confirmed the successful synthesis of Fe-Cu co-doped g-C3N4 with high purity, nano-sheet structure and high porosity (79.93 m2/g). The complete APAP decomposition efficiency was achieved under optimal experimental conditions including pH of 11, catalyst dosage of 10 mg/L, PS dosage of 1 mM, and APAP concentration of 4 mg/L. The scavenging tests confirmed the major contribution of sulfate radicals and consequently, hydroxyl radicals for APAP removal. In addition, the kinetics of APAP degradation was studied and it revealed the pseudo first–order kinetics with 0.0698 min−1 rate constant. Finally, a plausible and tentative decomposition pathway was proposed for APAP degradation. The results of this study confirmed that the LED/catalyst (Cu, Fe@g-C3N4)/PS process could be an efficient and robust process for antibiotic-containing wastewater including hospital wastewater

Effects of vitamin D supplementation on liver fibrogenic factors in non-alcoholic fatty liver patients with steatohepatitis: study protocol for a randomized clinical trial

Abstract Background It has been suggested that vitamin D and its receptors involve in suppressing fibrogenic signaling in non-alcoholic fatty liver disease (NAFLD). However, the effect of vitamin D supplementation on fibrogenic factors has not been investigated in NAFLD individuals with steatohepatitis. This study was designed to examine the effects on vitamin D supplementation on serum levels of vitamin D receptor (VDR), fibrogenic factors, and fibrogenic microRNAs (MiR) in NAFLD patients. Methods Forty-six NAFLD patients will be recruited in this study. After block matching for sex and BMI, they will be randomly assigned to receive 4000 IU/day vitamin D or placebo for 12 weeks. Weight, height, and waist circumference will be measured. Determination of serum fibrogenic MiRs, laminin, collagen type IV, hyaluronic acid, vitamin D, VDR, calcium, blood glucose, serum insulin, lipid profile, liver markers (ALT, AST, total, direct, and indirect bilirubin) will be done at study baseline and at the end of the trial. Insulin resistance and insulin sensitivity will be determined using the HOMA-IR and QUICKI equation. Discussion This is the first randomized controlled trial that will determine the effect of vitamin D supplementation on serum levels of VDR, fibrogenic factors, and fibrogenic MiRs in NAFLD patients. The results of this trial will provide clinical evidence on the effectiveness of vitamin D supplementation in controlling liver fibrosis in NAFLD patients. Trial registration Iranian Registry of Clinical Trials, IRCT201405251485N13. Registered on 14 March 2017

Prioritization of watersheds in order to perform administrative measures using fuzzy analytic hierarchy process

Prioritization of watersheds in order to perform administrative measures is necessary and inevitable. Determining areas of top priority for flood control projects is a managerial decision that should be approved by studies of physical, social and economic status of the region of interesrt and by assessing the outcomes of the past operations. Therefore, the aim of this research was to study morphological and physiographic characteristics, and to use geographic information systems (GIS) and multi-criteria decision-making methods (MCDM), to identify the critical sub-basins which have the tendency to be destructed, in Galikesh watershed, Golestan province. This watershed is important, yet critical, in terms of land use change, erosion and flooding in the Golestan Province, Iran. In total, nine morphological parameters were used to prioritize sub-watersheds using fuzzy analytic hierarchy process (FAHP). The morphological parameters were by some means linked to watershed drainage system. Based on FAHP approach, sub-basins, as vulnerable zones, have been evaluated and cetegorized in five priority levels (very low, low, medium, high and very high levels). The results showed that 44.44% and 22.22% of sub-basins were categorized respectively under average, and high to very high levels, suggesting that the conservation and management measures are essential in order to maintain stability in the region. Thus, the FAHP technique is a practical and convenient method to show potential zones in order to implement effective management strategies, especially in areas where data availability is low and soil diversity is high. Finally, it can be said that without having to encounter high costs and a waste of time, sub-basins could be categorized by means of morphometric parameters in order to implement conservational measures to simutaneously conserve soil and the environmen

A Nutrigenetic Approach to Investigate ApoB EcoR1 Polymorphism–Dietary Acid Load Interactions on Lipid and Anthropometric-Related Outcomes in Adults with Dyslipidemic Type 2 Diabetes

Introduction: Despite multiple studies which have considered the role of dietary acid load (DAL) or Apolipoprotein B (ApoB) EcoR1 polymorphism in diabetes, none have assessed their interplay effect on metabolic markers. Therefore, this study aimed to determine the interaction of EcoR1 and DAL on metabolic markers among adults with Type 2 diabetes mellitus (T2DM). Methods: 492 randomly selected individuals with T2DM were recruited for this cross-sectional study. Dietary intake was evaluated by a validated food frequency questionnaire. DAL was assessed as net-endogenous acid production (NEAP) and potential renal acid load (PRAL). Real-time-PCR was used to genotype EcoR1. Metabolic markers were also assessed. The interaction effect of the polymorphism and DAL indexes was analyzed by analysis of covariance (ANCOVA). Result: The frequency of EcoR1 genotypes was not different between dyslipidemic and normolipidemic participants (P>0.05). Among participants with dyslipidemia, those with the GG genotype and who consumed a higher level of NEAP had higher body mass index (BMI) (p=0.03) and waist circumference (WC; p =0.02). Moreover, triglyceride (TG) concentration (P=0.007), the LDL/HDL ratio (P=0.03) and the TG/HDL (P=0.03) ratio were significantly higher in A allele carriers with higher than the median intake of NEAP, in comparison with GG homozygotes. Finally, GA/AA carriers who had a higher intake of PRAL had a higher TG concentration (P=0.006) and TG/HDL ratio (P=0.01) compared to lower median intake in the dyslipidemia group. Discussion/Conclusion: In the dyslipidemic group, there was a higher TG concentration among individuals with the GA/AA genotype and a higher intake of NEAP/ PRAL. Also, in this group, a higher intake of NEAP may be considered as a risk factor for increased levels of BMI and WC among participants with the GG genotype

Dietary acid load and its interaction with CETP TaqB1 polymorphisms on lipid profile among patients with Type 2 diabetes mellitus

Abstract Objective Gene-diet interaction plays a key role in the inter-individual differences in lipid abnormalities as a major risk factor for cardiovascular diseases (CVDs). Thus, we explored the interaction between CETP TaqB1 polymorphism with dietary acid load (DAL) on lipid profile among type 2 diabetes mellitus (T2DM). Method This cross-sectional study conducted on 220 Iranian patients with T2DM. Dietary acid load (PRAL and NEAP) was calculated via a validated food-frequency questionnaire (FFQ). The polymerase chain reaction (PCR) used for genotyping Taq1B polymorphism. Biochemical markers were measured by standard protocol. The interaction between CETP Taq1B polymorphism and DAL (PRAL and NEAP) on lipid profile was performed by a generalized linear regression model (GLM). Results The overall prevalence of rs708272 genotypes was 8.6%, 72.7% and 18.6% for B1B1, B1B2 and B2B2 genotype respectively. This study showed that people with the B1B1 genotype had greater LDL, TC, LDL/HDL, and TG when they consumed diets that scored higher on the NEAP and PRAL indexes than those with the B1B2 and B2B2 genotypes. Besides, carriers of the B1B1 allele who were in the highest tertile of NEAP, had lower HDL (P Interaction < 0.05). Conclusions In summary, the lipid profile might be improved in B1B1 homozygotes by less adherence to DAL indexes, however, the findings should be validated in high-quality interventional studies