CYP2C19*17 association with higher plasma 4-hydroxy tamoxifen in Pakistani (estrogen-positive) breast cancer patients.

Breast cancer (BC) continues to be the most common cancer in the women worldwide. Since estrogen receptor (ER)-positive BC accounts for the majority of newly diagnosed cases, endocrine therapy is advised to utilize either tamoxifen (Tam) or aromatase inhibitors. The use of Tam as a monotherapy or in conjunction with an aromatase inhibitor following two or three years of endocrine therapy has long been recommended. When used adjuvantly, Tam medication reduces BC mortality and relapses, while it extends survival times in metastatic BC when used in conjunction with other treatments. Unfortunately, the efficiency of Tam varies considerably. This study was conducted to explore the influence of genetic polymorphisms in CYP2C19 gene on Tam's pharmacogenetics and pharmacokinetics in estrogen-positive BC patients. Data from healthy, unrelated individuals (n = 410; control group) and ER-positive BC patients (n = 430) receiving 20 mg of Tam per day were recruited. Steady-state plasma concentrations of Tam and its three metabolites were quantified using the high-performance liquid chromatography in the patients. The CYP2C19 polymorphisms were genotyped using an Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR) approach. More than 65% of healthy individuals were extensive metabolizers (*1/*1) for CYP2C19, whereas more than 70% of ER-positive BC patients were rapid and ultrarapid metabolizers (*1/17*, *17/17*). The polymorphism CYP2C19*17 is significantly associated with higher 4-hydroxytamoxifen (4-OH-Tam). Patients with the *17/*17 genotype exhibited 1- to 1.5-fold higher 4-OH-Tam, which was also high in patients with the *1/*2 and *2/*2 genotypes.

Estimating disparities in breast cancer screening programs towards mortality, case fatality, and DALYs across BRICS-plus.

BACKGROUND: Numerous studies over the past four decades have revealed that breast cancer screening (BCS) significantly reduces breast cancer (BC) mortality. However, in BRICS-plus countries, the association between BCS and BC case fatality and disability are unknown. This study examines the association of different BCS approaches with age-standardized mortality, case-fatality, and disability-adjusted life years (DALYs) rates, as well as with other biological and sociodemographic risk variables, across BRICS-plus from a national and economic perspective. METHODS: In this ecological study applying mixed-effect multilevel regression models, a country-specific dataset was analyzed by combining data from the Global Burden of Disease study 2019 on female age-standardized BC mortality, incidence, and DALYs rates with information on national/regional BCS availability (against no such program or only a pilot program) and BCS type (only self-breast examination (SBE) and/or clinical breast examination (CBE) [SBE/CBE] versus SBE/CBE with mammographic screening availability [MM and/or SBE/CBE] versus SBE/CBE/mammographic with digital mammography and/or ultrasound (US) [DMM/US and/or previous tests] in BRICS-plus countries. RESULTS: Compared to self/clinical breast examinations (SBE/CBE) across BRICS-plus, more complex BCS program availability was the most significant predictor of decreased mortality [MM and/or SBE/CBE: - 2.64, p < 0.001; DMM/US and/or previous tests: - 1.40, p < 0.001]. In the BRICS-plus, CVD presence, high BMI, second-hand smoke, and active smoking all contributed to an increase in BC mortality and DALY rate. High-income and middle-income regions in BRICS-plus had significantly lower age-standardized BC mortality, case-fatality, and DALYs rates than low-income regions when nationwide BC screening programs were implemented. CONCLUSIONS: The availability of mammography (digital or traditional) and BCS is associated with breast cancer burden in BRICS-plus countries, with regional variations. In light of high-quality evidence from previous causal studies, these findings further support the preventive role of mammography screening for BCS at the national level. Intervening on BCS related risk factors may further reduce the disease burden associated with BC.

Epidemiological and sociodemographic transitions of female breast cancer incidence, death, case fatality and DALYs in 21 world regions and globally, from 1990 to 2017: An Age-Period-Cohort Analysis.

Introduction: Breast cancer (BC) is the most widely studied disease due to its higher prevalence, heterogeneity and mortality. Objectives: This study aimed to compare female BC trends among 21 world regions and globally over 28 year of data and to assess the association between sociodemographic transitions and female BC risks. Methods: We used Global burden of disease study data and measure the female BC burden according to 21 world regions and sociodemographic indices (SDI). Age-period-cohort (APC) analysis was used to estimate time and cohort trend of BC in different SDI regions. Results: By world regions, age-standardised rate of female BC incidence were high in high-income-North America (ASR, 92.9; (95 %UI, 89.2, 96.6)), Western Europe (84.7; (73.4, 97.2)) and Australia (86; (81.7, 90.2)) in 2017. Whereas this rate was significantly increased by 89.5% between 1990 and 2017 in East Asia. We observed negative association between SDI and death, and DALYs in 25th and below percentiles of death and DALYs for the worldwide regions. Further, there was observed a strong negative correlation between SDI and case fatality percent (r2017 = -0.93; r1990 = -0.92) in both 2017 and 1990 worldwide and highest case fatality percentage was observed in Central Sub-Saharan Africa. Overall, the risk of case-fatality rate tends to decrease most noticeably in high middle SDI countries, and the reduction of the risk of case-fatality rate in the recent cohort was the lowest in the low SDI countries. Conclusions: Remarkable variations exist among various regions in BC burden. There is a need to reduce the health burden from BC in less developed and under developing countries, because under-developed countries are facing higher degree of health-related burden. Public health managers should execute more classified and cost-effective screening and treatment interferences to lessen the deaths caused by BC, predominantly among middle and low SDI countries having inadequate healthcare supplies.

Evaluation of lifestyle risk factor differences in global patterns of breast cancer mortality and DALYs during 1990-2017 using hierarchical age-period-cohort analysis.

BACKGROUND: Statistical evidence on breast cancer (BC) burden related to health and lifestyle risk factors are valuable for health policy-making. This study aimed to compare the trends in BC mortality and disability adjusted life years (DALYs) attributable to various health and life style risk factors among different world's regions according to sociodemographic index (SDI). METHODS: We extracted the age-standardized and age-specific rate of mortality and DALYs of women BC during 1990-2017 using the comparative risk assessment framework of the 2017 global burden of disease (GBD) study. We performed hierarchical age-period-cohort analysis to estimate age- and time-related trends, and effect of interactions between different risk factors on BC risk. RESULTS: During 1990-2017, the age-standardized rate of mortality and DALYs of women BC was increasing in less developed and under developing regions. The risk factor alcohol use [RR 51.3(95%CI 17.6-149.7)] and smoking [5.9(2.0-17.3)] were significantly highly contributor to increased mortality risk in high SDI region. Whereas in the low-SDI region, the greater mortality risk was observed in alcohol use [6.9(2.4-17)] and high FPG [2.7(1.5-3.1)]-related deaths. The adjusting for individual age, period, and risk factor effects, the significant interaction effect between metabolic risk factors and older ages were observed in all SDI regions and globally as well. However, an increasing cohort effect of alcohol, high fasting plasma glucose (FPG) and smoking-related death, and DALYs was observed during 1960 to 1985 cohorts among low-SDI regions. CONCLUSIONS: The age-standardized rates of mortality and DALYs due to BC has been increasing in low-SDI region. Alcohol consumption, high body mass index (BMI), high FPG, and smoking are potential BC risk factors particularly in older ages that leading to adverse disease outcomes. Therefore, rapid aging and prevalence of these prospective risk factors may strengthen the increasing mortality and DALYs of BC in low-SDI region. Hence, preventive measure along with strict action against concerned BC risk factors should be taken to reduce the disease burden specifically among lower-SDI regions.

Statistical analysis of COVID-19 infection caused by environmental factors: Evidence from Pakistan.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has become a severe public health problem around the globe. Various epidemiological, statistical, and laboratory-based studies have shown that the role of temperature and other environmental factors has important influence in the transmission of coronaviruses. Scientific research is needed to answer the questions about the spread and transmission of the infection, whether people could be avoided from being infected with COVID-19 in next summer. AIM: We aim to investigate the association of daily average temperature, daily average dew point, daily average humidity, daily average wind speed, and daily average pressure with the infection caused by this novel coronavirus in Pakistan. KEY FINDINGS: First, we check the correlation between environmental factors and daily infected cases of COVID-19; among them, temperature and dew point have positive linear relationship with daily infected cases of COVID-19. The thought-provoking findings of the present study suggested that higher temperature and dew point can contribute to a rise in COVID-19 disease in four provinces of Pakistan, possible to genome modifications and viral resistance to harsh environment. Moreover, it is also observed that humidity in Punjab and Sindh, and wind speed in Balochistan and Khyber Pakhtunkhwa have influenced the spreading of daily infected COVID-19 cases. SIGNIFICANCE: Current study will serve as a guideline to develop understanding of environmental factors that influence COVID-19 spread, helping policymakers to prepare and handle a catastrophe resulting from this pandemic.

Correlation of MSH2 exonic deletions and protein downregulation with breast cancer biomarkers and outcome in Pakistani women/patients.

Mismatch repair (MMR) pathway is one of the underlying mechanisms of predisposition to breast cancer (BC). The present study explored the association of MSH2 exonic deletions, respective survival analysis, protein structure prediction, transcription profiling, and expression analysis with BC risk. Genotyping analysis of 493 BC cases and 387 controls confirmed the association of two MSH2 exonic deletions, i.e., exon 3 (OR:6.4, CI = 3.4-12.1) and 9 (OR:7.8, CI = 4.1-14.8) with BC risk. In order to confirm the phenotypic-genotypic relationship, we have performed MSH2 transcriptomic (p < 0.05) and protein expression analysis (OR:30, CI = 4-230) which further confirmed its downregulation/loss in BC biopsy samples highlighting potential role in the onset of breast carcinogenesis. Additionally, we have presented that MSH2 mutations can alter the expression profile of other BC associated biomarkers like ER, PR, CK-7, GATA-3, and E-cadherin. Subsequently, the effect of exonic deletions on secondary structure of protein has shown missing of beta and alpha helices in their protein products via in-silico analysis. However, loss of exon 3 results in the altered core protein structure leading to dysfunction protein, possible cause of BC development. No association of MSH2 exonic deletions with survival statistics was observed conceivably due to the shorter follow-up time. Thus, our results at genetic, transcriptomic, and proteomic levels confirmed the downregulated MSH2, emphasizing its potential contribution in MMR mechanisms for breast tumorigenesis. In conclusion, MSH2 deficiency may cause breast cancer development and progression.

Possible Potential Outcomes from COVID-19 Complications on Testes: Lesson from SARS Infection.

Coronavirus infection disease-2019 (COVID-19) gained worldwide fame after deadly outbreak in China and its subsequent spread to many countries. So far, COVID-19 is not fully contained, and new cases are arising on daily bases in various countries. Due to zoonotic nature and human-to-human spread, COVID-19 is considered pandemic with more causalities in developing countries. Full genome analysis revealed its resemblance with severe acute respiratory syndrome (SARS) virus with minor variation in non-structural proteins. Both viruses use the common angiotensin converting enzyme (ACE2) receptor to attach to the epithelial cells of the target organs. Currently, COVID-19 patients are treated with drugs that are used for lungs infections. However, ACE2 has high expression in other human organs such as kidney and testes. Thus, it is assumed that, like SARS, it may have adverse effects on other vital organs, which have dominant expression of ACE2. In testis, SARS affected patients displayed peritubular fibrosis, extensive germ cell disruption, damage of blood-testis barrier integrity and more frequent occurrence of apoptosis. Here, we critically discuss the possible adverse effects of COVID-19 on the testes of patients along with future precautions to overcome the complications of reproductive organs. Key Words: COVID-19; SARS; ACE2, Testes.

Unraveling the Epidemiology, Geographical Distribution, and Genomic Evolution of Potentially Lethal Coronaviruses (SARS, MERS, and SARS CoV-2).

SARS CoV appeared in 2003 in China, transmitted from bats to humans via eating infected animals. It affected 8,096 humans with a death rate of 11% affecting 21 countries. The receptor binding domain (RBD) in S protein of this virus gets attached with the ACE2 receptors present on human cells. MERS CoV was first reported in 2012 in Middle East, originated from bat and transmitted to humans through camels. MERS CoV has a fatality rate of 35% and last case reported was in 2017 making a total of 1,879 cases worldwide. DPP4 expressed on human cells is the main attaching site for RBD in S protein of MERS CoV. Folding of RBD plays a crucial role in its pathogenesis. Virus causing COVID-19 was named as SARS CoV-2 due its homology with SARS CoV that emerged in 2003. It has become a pandemic affecting nearly 200 countries in just 3 months' time with a death rate of 2-3% currently. The new virus is fast spreading, but it utilizes the same RBD and ACE2 receptors along with furin present in human cells. The lessons learned from the SARS and MERS epidemics are the best social weapons to face and fight against this novel global threat.

Trend Dynamics of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Transmission in 16 Cities of Hubei Province, China.

OBJECTIVE: A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was detected by researchers from a patient in Wuhan, Hubei province, China, in December 2019, and broke out in January 2020. Then, the pandemic was detected in countries around the world. Therefore, precise estimates of its current and future trends are highly required for future policy implications. METHODS: We retrieved data from the Health Commission of Hubei, China. Logistic-S curve model was used to estimate the current and future trends of SARS-CoV-2-infected cases among 16 cities of Hubei, China from Jan-11 to Feb-24, 2020. RESULTS: Out of 64,287 confirmed cases of SARS-CoV-2 infection in Hubei, higher percentage of cases were in Wuhan and Xiaogan. The highest death percentage was found in Wuhan and Qianjiang. A significant percentage of cures were found in Enshi Prefecture and Huanggang, while Wuhan showed the lowest percentage of cures. Rising trends in infected cases were observed throughout the study period, particularly in Wuhan, and a higher trend was observed after 12-Feb. Gradual decline trend of SARS-CoV-2 cases was observed during Feb-25 to Mar-15 in Hubei Province. Future forecast showed that the average number of SARS-CoV-2-infected cases might be decreased or stable in Hubei in the coming 20 days. CONCLUSION: The public must take precautionary measures in order to control and prevent disease spread and avoid extra travelling.

XPC as breast cancer susceptibility gene: evidence from genetic profiling, statistical inferences and protein structural analysis.

BACKGROUND: Gene polymorphisms that affect nucleotide excision repair (NER) pathway may link with higher susceptibility of breast cancer (BC); however, the significance of these associations may vary conferring to the individual ethnicity. Xeroderma pigmentosum complementation gene (XPC) plays a substantial role in recognizing damaged DNA during NER process. OBJECTIVE AND METHODS: To estimate the relationship among XPC polymorphisms and breast cancer (BC) risk, we carried out a case-control-association study with 493 BC cases and 387 controls using TETRA-ARMS-PCR. Distributional differences of clinical features, demographic factors and XPC polymorphisms among BC cases and controls were examined by conditional logistic regression model. Kaplan-Meier test was applied to predict survival distributions and protein structure was predicted using computational tools. RESULTS: Obesity, consanguinity, positive marital status and BC family history were associated (P ≤ 0.01) with higher BC risk. Genotyping revealed significant involvement (P ≤ 0.01) of two XPC polymorphisms rs2228001-A > C (OR = 3.8; CI 1.9-7.6) and rs2733532-C > T (OR = 2.6; CI 1.4-5.03) in BC development, asserting them potential risk factors for increased BC incidence. However, no association (P > 0.05) was detected for overall or progression free survival for both XPC polymorphisms possibly due to shorter follow-up time (45 months). As compared to normal XPC structure, pronounced conformational changes have been observed in the C-terminus of XPCQ939K, bearing rs2228001-A > C substitution. In XPCQ939K, two additional α-helices were observed at A292-E297 and Y252-R286, while L623-M630 and L649-L653 helices were converted into loop conformation. CONCLUSION: In conclusion, both XPC polymorphisms confer significant association with increased BC risk. rs2228001 substitution may change the structural and functional preferences of XPC C-terminus, while rs2733532 may have regulatory role thereby leading to potential BC risk.

A Hierarchical Age-Period-Cohort Analysis of Breast Cancer Mortality and Disability Adjusted Life Years (1990-2015) Attributable to Modified Risk Factors among Chinese Women.

Limited studies quantified the age, period, and cohort effects attributable to different risk factors on mortality rates (MRs) and disability-adjusted life years (DALYs) due to breast cancer among Chinese women. We used data from the Global Burden of Disease Study (GBD) in 2017. Mixed-effect and hierarchical age-period-cohort (HAPC) models were used to assess explicit and implicit fluctuations in MRs and DALYs attributable to different breast cancer associated risk factors. As the only risk factor, high body mass index (HBMI) showed continuously increasing trends in MRs and DALYs across ages, periods, and cohorts. Age, recent periods (2010-2015), and risk factor HBMI showed significant positive effect on MRs and DALYs (p < 0.05). Moreover, we reported significant interaction effects of older age and period in recent years in addition to the interplay of older age and risk factor HBMI on MRs and DALYs. Increased age and obesity contribute to substantially raised breast cancer MRs and DALYs in China and around the globe. These discoveries shed light on protective health policies and provision of healthy lifestyle for improving the subsequent breast cancer morbidity and mortality for China, as well as other related Asian regions that are presently facing the same public health challenges.

Diagnostic accuracy of cannabinoid testing by liquid chromatography-tandem mass spectrometry in human hair.

OBJECTIVE: To determine the diagnostic accuracy of Cannabinoids testing by LC-MS/MS in human hair and compare it with urine in civil heavy vehicle drivers. METHODS: Current study was a diagnostic accuracy study done in "Armed Forces Institute of Pathology Rawalpindi, Pakistan" from February to November 2017. Urine and hair samples were collected by non-probability convenient sampling technique from 151 heavy vehicle drivers from Punjab. Hair and urine samples were collected from each subject. Separation of compounds was performed on Agilent Poroshell and analyzed using 6460 Triple Quadrapole LC-MS along-with software Mass hunter ©. RESULTS: Study population (151 civil heavy vehicle drivers) was divided into three main divisions There were 69 (46%) truck drivers,43 (28.5%) twenty-wheeler drivers and 39 (26%) bus drivers. Mean age of study participants was 36±10.82 years. Paired t-test was applied to check mean difference between the two tests' concentration (i.e urine and hair analysis for cannabis) which showed significant difference at p<0.001. Among the different factors of diagnostic accuracy in hair and urine specimens were: Sensitivity (96% and 62%), Specificity (93% and 95%) Positive Predictive Value (88% and 87%), Negative Predictive Value (97% and 82%) respectively. Overall diagnostic accuracy of Cannabinoids detection in hair was 94% while in urine it was 83%. ROC curve showed area under curve of 0.79 and 0.96 for urine and hair samples respectively. CONCLUSIONS: Current study signified hair as a substitute matrix owing to its non-invasive specimen collection, better diagnostic yield and wider detection period compared to urine.

Correlation of MLH1 polymorphisms, survival statistics, in silico assessment and gene downregulation with clinical outcomes among breast cancer cases.

This study aimed to investigate the role of MLH1 polymorphisms, respective protein structure prediction, survival analysis, related clinicopathological details and MLH1 expression in breast cancer (BC). Genotyping of selected SNPs in BC patients (493) and age matched controls (387) were performed by Tetra-ARMS PCR. Gene expression among breast tumors (127) and adjacent control tissues were analysed using reverse transcriptase PCR (RT-PCR) and immunohistochemistry. Statistical analysis was performed by SPSS and MedCalc. Conditional logistic regression analysis was applied to compute the odds ratio and confidence interval. Phyre2 and I-TASSER were used to generate MLH1 protein structures and verified by a variety of computational tools. Genotyping illustrated that MLH1 polymorphisms (rs63749795 and rs63749820) were significantly associated (P ≤ 0.05) with risk of developing BC. Down regulation of MLH1 gene expression/loss of the MLH1 protein (OR 12; CI 2.8-53.1) was observed in BC cases, illustrating its potential role in disease development. Moreover, loss of the MLH1 protein was found to be associated with higher grade cancer (P = 0.02) and lymph node positivity (P = 0.03), highlighting its essential role, as a component of the mismatch repair (MMR) machinery. Bioinformatics analysis confirmed that nonsense mutations produce a truncated MLH1 protein, causing a reduction in MMR efficiency. No association between MLH1 polymorphisms and overall and progression free survival statistics was observed among BC cases, possibly due to short follow-up study. Results at DNA, RNA and protein levels, along with in silico analysis, highlights the potential role of MLH1 in DNA repair mechanisms, within BC. Therefore, it was concluded that MLH1 may contribute towards BC development and progression.

Recent insights into breast cancer incidence trends among four Asian countries using age-period-cohort model.

PURPOSE: Breast cancer is one of the rapidly increasing cancers among women and a significant cause of cancer-related morbidity and mortality worldwide. Therefore, the current study was designed to examine and compare trends of breast cancer incidence (BCI) during the observed period (1990-2015) in specific age groups and investigate age-specific, time period, and birth cohort-related effects on BCI in China, India, Pakistan, and Thailand. PATIENTS AND METHOD: Data related to BCI were retrieved from the Institute for Health Metrics and Evaluation. Age-period-cohort model joint with intrinsic estimator algorithm was used to estimate the effect of age, period, and birth cohort on BCI. BCI rates were analyzed among different age groups ranging from 20 to 84 years in specified periods. RESULT: Overall, results showed an increasing trend of BCI among four Asian countries during the study period especially in age groups 50 to 84 years. Higher incidence rates were observed in 2015 in the age group 70-74, 65-69, 50-54, and 60-64 in Pakistan, China, India, and Thailand, respectively. Age period cohort analysis revealed significantly raised effect of age and period and declined effect of the cohort on incidence rates. CONCLUSION: The current study reported increased BCI with time in selected four Asian countries. Overall, BCI remained high in Pakistan as compared to China, India, and Thailand. Although proper registries are not available in most of the developing Asian countries, the current study highlighted the increased incidence and may play an essential role in registries development or spreading awareness against this disease. Therefore, maintaining proper records to build registries at the national level along with advancements in breast cancer screening and treatment are highly recommended to deal with the increasing burden of this disease.

Hypertension associated risk factors in Pakistan: A multifactorial case-control study.

OBJECTIVE: To shed light on the association of age, smoking, educational status, family history, diabetes and kidney diseases with hypertension. METHODS: The case-control study was conducted at three different medical centres of Rawalpindi, Pakistan, from December 2016 to July 2017. Data from 549 hypertensive cases and 1451 normotensive controls was collected using a detailed questionnaire and through personal interviews by adopting nonprobability consecutive sampling technique. Overall 2000 adult individuals, both males and females excluding pregnant women, were the part of this study. Those with blood pressure ≥140/90mmHg and taking anti-hypertensive treatment were designated as the cases, while the rest were taken as normotensive controls. Blood pressure was measured by a physician. Multivariate logistics regression analysis was used to estimate the association of various different risk factors with hypertension. All the analysis was performed using software R 3.4.2 and SPSS 24. RESULTS: Of the 2,000 subjects, 549(27.45%) were hypertensive cases and 1451(72.55%) were normotensive controls. Mean age of the cases was 43.32}9.7 years and it was 31.8}10.1 years among the normotensives. Higher age, smoking, lower educational status, presence of kidney diseases, diabetes and family history of hypertension were significantly associated with hypertension (p<0.01 each). CONCLUSIONS: In Pakistani population, age, smoking, illiteracy, kidney diseases, diabetes and family history were found to be associated with hypertension.

Survival analysis of breast cancer patients with different treatments: a multicentric clinicopathological study.

OBJECTIVE: To explore and better understand clinic pathological details of breast cancer patients and analyse their survival rate among different treatment groups. METHODS: The prospective cohort, multi-centric study was conducted from September, 2014, to February, 2018, at five hospitals in Rawalpindi and Islamabad, Pakistan, and comprised histo-pathologically confirmed breast cancer cases. Patient characteristics and medical history were collected using a detailed questionnaire. All the subjects were followed up, and information regarding their current health and treatment status was collected. Data was analysed using SPSS 24. RESULTS: There were 347 subjects with a mean age of 44.3±12.2 years and body mass index of 27.9±4.0 kg/m2. Younger age, increased body mass index, consanguinity and family history were major contributing factors in breast cancer development (p<0.05). Overall, 267(77%) had invasive ductal carcinoma and Grade II tumour 234(67%) was more frequent. A total of 221(64%) cases had positive lymph nodes and 97(28%) had metastasis to different body organs. Overall survival analysis showed statistically significant role (p<0.0001) of all treatment options. CONCLUSIONS: Combination of different treatments can provide more promising health outcomes in breast cancer cases.

Expressional analysis of MLH1 and MSH2 in breast cancer.

BACKGROUND: Mismatch repair proteins are ubiquitous keys in diverse cellular functions and protects the genome by correcting mismatch as post replication error correction machinery. Mismatch repair deficiency was associated with tumor development and progression therefore, current study was aimed to investigate MLH1 and MSH2 expression in breast cancer and correlate patients' clinicopathological factors with status of mismatch repair genes. MATERIAL AND METHODS: Breast cancer tissues with adjacent normal tissue along with clinical details were collected during surgery from 80 cases. Immunohistochemistry was performed with primary and secondary antibodies for expressional analysis. Results were analyzed using SPSS version 24. RESULTS: Immunohistochemical analysis revealed that both MLH1 and MSH2 were crucial in maintaining DNA repair system and loss of these 2 mismatch repair proteins may lead to adverse outcomes in breast cancer. Statistically significant association was found between loss of MLH1 (P = 0.0004; odds ratio 13.8; 95% confidence interval 4.6-41.1), MSH2 (P = 0.0002; odds ratio 14.0; 95% confidence interval 4.7-42.2) and breast cancer. Statistical analysis demonstrated that MLH1 and MSH2 deficiency may lead breast cancer progression to advanced stage, correlated with tumor focality (MLH1 P = 0.001; MSH2 P = 0.002) and chemotherapy (MLH1 P = 0.01; MSH2 P = 0.04). Presence of CK7, GATA 3, and E cadherin tends to increase in mismatch repair deficient breast cancer. Whereas, no association of mismatch repair deficiency was observed with age, tumor grade, positive lymph nodes, menopause, and ER and/or PR status. CONCLUSION: Loss of mismatch repair proteins in breast cancer highlights its potential role in DNA repair mechanisms and helps tumor cells to become resistant against chemotherapeutic drugs. Therefore, mismatch repair deficiency may contribute to breast cancer progression.

Determination of reference intervals of thyroid markers during pregnancy in Urban area of District Rawalpindi Pakistan.

OBJECTIVE: To determine the reference values for thyroid stimulating hormone, free tetra-iodothyronine and total tri-iodothyronine for healthy pregnant women. METHODS: This cross sectional study was conducted at the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from January 2016 to June 2017. Pregnant women with normal, single intrauterine, uncomplicated pregnancy were recruited from the local population. Blood sample was taken to analyse thyroid stimulating hormone, free tetra-iodothyronine and total tri-iodothyronine using chemiluminescence immunoassay. For thyroid hormone levels during each trimester 5th and 95th percentiles were calculated as reference intervals. Data was analysed using SPSS 24. RESULTS: Out of 384 subjects, 188(48.95%) were in their first trimester and 196(51.04 %) females were in their second trimester. There were 109(57.97%) primigravida in the first trimester and 137(69.9%) in the second trimester. Mean age of subjects presenting in the first and second trimester was 25.37±3.78 years and 26.54±4.65 years respectively. Reference intervals for those in the first trimester for thyroid stimulating hormone was 0.05-2.8uIU/ml, for free tetra-iodothyronine14.4-22.7pmol/l and total tri-iodothyronine1.5-3.3nmol/l. For those in second trimester the corresponding values were 0.16-3.3 uIU/ml, 14.2-24.6.0 pmol/l and 1.6-3.1nmol/l. CONCLUSIONS: Laboratories should adopt trimester-specific reference intervals for thyroid function tests in pregnancy..

An insight into clinical outcome of XPG polymorphisms in breast cancer.

XPG polymorphisms are associated with varied clinical outcomes in different cancers but up-till now no study has been reported on breast cancer. Therefore, current study was aimed to explore the association of breast cancer risk factors and XPG polymorphisms (rs2296147 and rs1047768). It also investigated impact of XPG variants on overall survival and progression free survival among breast cancer cases. A total of 493 histopathologically identified breast cancer cases and 387 healthy females were genotyped by ARMS-PCR. Relationship between general characteristics, XPG polymorphisms and breast cancer risk was accessed by conditional logistic regression and illustrated by OR and 95% CI. Kaplan Meier test was applied to estimate survival distributions whereas log rank test demonstrated survival differences. Association of XPG variants with OS and PFS in breast cancer was illustrated by HR and 95% CI. Early onset of menopause, consanguinity and family history contributed (P < 0.05) towards breast cancer development. Both rs2296147 and rs1047768 SNPs were found to be associated (P < 0.05) with the risk of breast cancer. XPG rs1047768 was significantly associated with decreased PFS (HR 1.72; 95% CI 1.0-2.8) in breast cancer cases (P = 0.013) which was demonstrated by median time of 26 months for T > C variant when compared with median time of 37 months for TT genotype. No association was found between XPG rs2296147 polymorphism and survival analysis among breast cancer cases. XPG (rs1047768 T > C) variant may play a significant role in terms of decreased PFS and could be used as a predictor of unfavourable prognosis among breast cancer.