Validation of Self-Reported Smoker and Second Hand Smoke Exposure by Urinary Cotinine within The Malaysian Cohort Project

Background: Validation of self-reported questionnaire is very crucial in ensuring the quality and reliability of data collection. Objective: The aim of this study were i) to validate the questionnaire on tobacco smoke intake and second hand smoke exposure among The Malaysian Cohort (TMC) subjects through the determination of urinary cotinine levels, ii) to determine the optimal cut-off point of urine cotinine that discriminates smokers from non-smokers and iii) to estimate misclassification rate between self-reported smoking and urinary cotinine level.Methods: Urine samples from a total of 775 The Malaysian Cohort subjects (104 smokers, 102 former smokers and 569 non-smokers) were obtained and urinary cotinine levels were determined by high-performance liquid chromatography (HPLC). Differences between groups were compared using Kruskal Wallis and Mann-Whitney tests. The Receiver Operating Characteristic (ROC) curved was performed to define the optimal urinary cotinine cut-off point.Results: Urinary cotinine concentration significantly (p<0.001) correlated with smoking status (r=0.46), the average number of cigarettes smoked per day (r=0.53), duration of smoking (r=0.33) and number of cigarettes packed per year (r=0.47). Smokers and second hand smokers have significantly higher median cotinine levels (978.40 and 21.31 respectively) compared to non-smokers (15.52) and non-exposed (13.60) subjects. Cotinine level at cut-off value of 1.51 ng/mg creatinine is able to distinguish smokers and non-smokers with a sensitivity of 84.62% and specificity of 81.97%.Conclusion: The Malaysian Cohort self-reported smoking questionnaire is a reliable tool in assessing the use of tobacco and second hand smoke exposure among the subjects

Expression of p16 and pKi-67 in cervical preneoplasia and neoplasia

This study was conducted to investigate the expression of p16 and pKi-67 in normal, preneoplasia and neoplasia lesions of the uterine cervix. One hundred and thirty one cervical specimens, consisting of normal cervix (n = 43), cervical intraepithelial neoplasia (CIN) lesions (n = 40) [16 LSIL: CIN 1 and 24 HSIL: 9 CIN 2 and 15 CIN 3] and cervical squamous cell carcinomas (n = 48) [16 SCC I, 17 SCC II, 7 SCC III and 8 SCC IV] were examined immunohistochemically in paraffin sections. All samples of the normal cervix were negative for p16. Immunoreactivity of p16 was observed in 4/ 16 LSIL, 12/24 HSIL and 30/48 SCC. In all p16-positive samples, both nuclear and cytoplasmic staining were observed. High expression of p16 (> 50 % of cell stained) was found in HSIL and SCC. Ki-67 index was significantly (p < 0.05) higher in high-grade squamous intraepithelial lesions (HSIL: CIN 2 & 3) and SCC lesions when compared to normal cervices. The expression of p16 and Ki-67 proliferation profile ( 30 % stained cells) were significantly associated with the grade of lesions (χ2 = 6.832, p = 0.033 and χ2 = 10.952; p = 0.012 respectively). There was no significant relationship demonstrated between p16 positivity and Ki-67 proliferation profile (χ2 = 0.292; p = 0.589). Our results indicated that p16protein may be involved in the carcinogenesis of cervical cancer. However, overexpression of p16 seemed to have no effect on cell proliferation. The expression of p16 and pKi-67 may be useful in cases where it is difficult to make a diagnosis by histology

Positive TPMT genotype-phenotype correlation underscores importance of TPMT genotyping for personalized thiopurine dosing

This study explored TPMT genotype-phenotype correlation in a group of acute lymphoblastic leukemia (ALL) patients to investigate the potential of TPMT genotyping for personalized thiopurine dosing. Genotyping for G238C (TPMT*2), G460A (TPMT*3B) and A719G (TPMT*3C) loci was determined in 89 subjects via PCR, while TPMT activity was measured using HPLC. TPMT*3C was the only mutant allele detected in 4 heterozygous carriers. These patients had significantly lower (23.0 nmol/g Hb/h) TPMT activity compared to wildtype patients (51.0 nmol/g Hb/h) (p = 0.003). Positive correlation between TPMT genotype and phenotype projects the possibility of using TPMT genotyping as a guide prior thiopurine drug administration.

Activated ADI pathway: the initiator of intermediate vancomycin resistance in Staphylococcus aureus

Comparative proteomic profiling between two vancomycin intermediate Staphylococcus aureus (VISA) strains, Mu50Ω-vraSm and Mu50Ω-vraSm-graRm, and vancomycin-susceptible S. aureus (VSSA) strain Mu50Ω, revealed up-regulated levels of catabolic ornithine carbamoyltransferase (ArcB) of the arginine catabolism pathway in VISA. Subsequent analyses showed that the VISA strains have higher levels of cellular ATP and ammonia, which are by-products of arginine catabolism, and displayed thicker cell walls. We postulate that elevated cytoplasmic ammonia and ATP molecules, resulting from activated arginine catabolism upon acquisition of vraS and graR mutations, are important requirements facilitating cell wall biosynthesis, thereby contributing to thickened cell wall and consequently reduced vancomycin susceptibility in VISA.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Chlorella vulgaris triggers apoptosis in hepatocarcinogenesis-induced rats*

Chlorella vulgaris (CV) has been reported to have antioxidant and anticancer properties. We evaluated the effect of CV on apoptotic regulator protein expression in liver cancer-induced rats. Male Wistar rats (200~250 g) were divided into eight groups: control group (normal diet), CDE group (choline deficient diet supplemented with ethionine in drinking water to induce hepatocarcinogenesis), CV groups with three different doses of CV (50, 150, and 300 mg/kg body weight), and CDE groups treated with different doses of CV (50, 150, and 300 mg/kg body weight). Rats were sacrificed at various weeks and liver tissues were embedded in paraffin blocks for immunohistochemistry studies. CV, at increasing doses, decreased the expression of anti-apoptotic protein, Bcl-2, but increased the expression of pro-apoptotic protein, caspase 8, in CDE rats, which was correlated with decreased hepatoctyes proliferation and increased apoptosis as determined by bromodeoxy-uridine (BrdU) labeling and terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) assay, respectively. Our study shows that CV has definite chemopreventive effect by inducing apoptosis via decreasing the expression of Bcl-2 and increasing the expression of caspase 8 in hepatocarcinogenesis-induced rats

36-month clinical outcomes of patients with venous thromboembolism: GARFIELD-VTE

Background: Venous thromboembolism (VTE), encompassing both deep vein thrombosis (DVT) and pulmonary embolism (PE), is a leading cause of morbidity and mortality worldwide.Methods: GARFIELD-VTE is a prospective, non-interventional observational study of real-world treatment practices. We aimed to capture the 36-month clinical outcomes of 10,679 patients with objectively confirmed VTE enrolled between May 2014 and January 2017 from 415 sites in 28 countries.Findings: A total of 6582 (61.6 %) patients had DVT alone, 4097 (38.4 %) had PE +/- DVT. At baseline, 98.1 % of patients received anticoagulation (AC) with or without other modalities of therapy. The proportion of patients on AC therapy decreased over time: 87.6 % at 3 months, 73.0 % at 6 months, 54.2 % at 12 months and 42.0 % at 36 months. At 12-months follow-up, the incidences (95 % confidence interval [CI]) of all-cause mortality, recurrent VTE and major bleeding were 6.5 (7.0-8.1), 5.4 (4.9-5.9) and 2.7 (2.4-3.0) per 100 person-years, respectively. At 36-months, these decreased to 4.4 (4.2-4.7), 3.5 (3.2-2.7) and 1.4 (1.3-1.6) per 100 person-years, respectively. Over 36-months, the rate of all-cause mortality and major bleeds were highest in patients treated with parenteral therapy (PAR) versus oral anti-coagulants (OAC) and no OAC, and the rate of recurrent VTE was highest in patients on no OAC versus those on PAR and OAC. The most frequent cause of death after 36-month follow-up was cancer (n = 565, 48.6 %), followed by cardiac (n = 94, 8.1 %), and VTE (n = 38, 3.2 %). Most recurrent VTE events were DVT alone (n = 564, 63.3 %), with the remainder PE, (n = 236, 27.3 %), or PE in combination with DVT (n = 63, 7.3 %).Interpretation: GARFIELD-VTE provides a global perspective of anticoagulation patterns and highlights the accumulation of events within the first 12 months after diagnosis. These findings may help identify treatment gaps for subsequent interventions to improve patient outcomes in this patient population