Urea cycle promotion via ammonia-upregulated CPS1 is involved in arsenite-induced pulmonary fibrosis through enhancing collagen synthesis.

Arsenic exposure is connected with lung toxicity and is related to lung fibrotic changes. Idiopathic pulmonary fibrosis (IPF) is characterized by extracellular matrix (ECM) deposition. Various genetic mechanisms and environmental factors induce or exacerbate pulmonary fibrosis. Collagen synthesis induced by sodium arsenite (NaAsO2) is closely associated with IPF. Fibroblasts tend to fine-tune their metabolic networks to support their synthetic requirements in response to environmental stimuli. Alterations in metabolism have an influential role in the pathogenesis of IPF. However, it is unclear how arsenic affects the metabolism in IPF. The urea cycle (UC) is needed for collagen formation, which provides adequate levels of proline (Pro) for biosynthesis of collagen. Carbamoyl phosphate synthetase 1 (CPS1) converts the ammonia to carbamoyl phosphate, which controls the first reaction of the UC. We show that, in arsenite-exposed mice, high amounts of ammonia in the lung microenvironment promotes the expression levels of CPS1 and the Pro metabolism. Reduction of ammonia and CPS1 ablation inhibit collagen synthesis and ameliorate IPF phenotypes induced by arsenite. This work takes advantage of multi-omics data to enhance understanding of the underlying pathogenic mechanisms, the key molecules and the complicated cellular responses to this pollutant, which provide a target for the prevention of pulmonary fibrosis caused by arsenic.

Intrauterine arsenic exposure induces glucose metabolism disorders in adult offspring by targeting TET2-mediated DNA hydroxymethylation reprogramming of HNF4α in developing livers, an effect alleviated by ascorbic acid.

Exposure to arsenic during gestation has lasting health-related effects on the developing fetus, including an increase in the risk of metabolic disease later in life. Epigenetics is a potential mechanism involved in this process. Ten-eleven translocation 2 (TET2) has been widely considered as a transferase of 5-hydroxymethylcytosine (5hmC). Here, mice were exposed, via drinking water, to arsenic or arsenic combined with ascorbic acid (AA) during gestation. For adult offspring, intrauterine arsenic exposure exhibited disorders of glucose metabolism, which are associated with DNA hydroxymethylation reprogramming of hepatic nuclear factor 4 alpha (HNF4α). Further molecular structure analysis, by SEC-UV-DAD, SEC-ICP-MS, verified that arsenic binds to the cysteine domain of TET2. Mechanistically, arsenic reduces the stability of TET2 by binding to it, resulting in the decrease of 5hmC levels in Hnf4α and subsequently inhibiting its expression. This leads to the disorders of expression of its downstream key glucose metabolism genes. Supplementation with AA blocked the reduction of TET2 and normalized the 5hmC levels of Hnf4α, thus alleviating the glucose metabolism disorders. Our study provides targets and methods for the prevention of offspring glucose metabolism abnormalities caused by intrauterine arsenic exposure.

Relationship between aging and control of metabolic syndrome with telomere shortening: a cross-sectional study.

Aging is considered one of the major risk factors for several human disorders. The telomere plays a crucial role in regulating cellular responsiveness to stress and growth stimuli as well as maintaining the integrity of the Deoxyribonucleic Acid (DNA), and aging leads to the progressive decline in the telomere length (TL) due to continuous cell division. The aim of this study was to determine the relationship between TL and advancing age and the impact of metabolic syndrome (MetS) on TL. Firstly, we determined the association of advancing age and TL, by measuring telomere length (T/S ratio) in healthy volunteers (n = 90). The TL was compared between normal population and patients with metabolic syndrome (n = 298). The age matched controlled and uncontrolled MetS patients (n = 149) were also compared for their TL T/S ratio. The TL showed negative correlation with advancing age, whereas the significant change was observed at the cut-offs of 40 and 70 years defining 40 with longer TL and 70 as shorter TL. The longest T/S ratio at 2.46 was measured at the age range of 1 year in healthy volunteers, while elderly population showed considerably shorter TL. The patients older than 60 years with poor or uncontrolled MetS had shorter TL, as compared to the controlled MetS. In conclusion our findings suggest that TL was negatively correlated with advancing age. Uncontrolled metabolic syndrome appeared to have worsening effects on TL. Telomere length appears to have potential to be used a parameter to determine age. However, further large scale studies are recommended to make firm guidelines.

Novel molecular classification of colorectal cancer and correlation with survival.

Introduction: Colorectal cancer (CRC) is one of the most common cancers worldwide. This study was designed to evaluate biological patterns, explore molecular classification and correlate with survival outcome in treatment naïve CRC patients. Methods: Over 11 years consecutive series of 435 CRC patients were operated on as primary surgical therapy. A total of 201 CRC patients were included, whose complete set of clinical information was available, and their good quality tumour blocks were retrieved. Immunohistochemistry was used for tumour analysis, and partitional clustering was performed using R software for cluster analysis. Results: The median age was 43 (range 10-85) years; adenocarcinoma was the most commonly seen histological type. The great majority had positive CK20, CEA, E-Cadherin, Ki67, CDX2, and p53 expression. There were four distinct molecular classes found, whereas Ki67, CDX2, and p53 play the main role in partitioning. Younger age negatively impacted survival; overall and disease-specific survival was 26 months only with 50 months' longest survival. Conclusion: Colorectal cancer is a biologically heterogeneous disease with at least four distinct molecular patterns, where cell proliferation and gene repair mechanisms appear to play the key role.

Association of Equiliberative Nucleoside Transporter (ENT) with liver fibrosis stage in Hepatitis-C.

OBJECTIVE: To evaluate the pattern of equiliberative nucleoside transporters in viral hepatitis responding and non-responding patients, to correlate the liver fibrosis stage with the pattern among the non-responders, and to correlate the equiliberative nucleoside transporter status with housekeeping hypoxanthine-guanine phosphoribosyltransferase gene. Method: The comparative cross-sectional study was conducted at the Molecular Biology and Genetics Department, Liaquat University of Medical and Health Sciences, Jamshoro, Pakistan, from March to August 2018, and comprised adult hepatitis C virus patients of either gender who completed six months of treatment. They were assessed for response to therapy in terms of the presence of the viral load in their serum by using real time polymerase chain reaction, and divided into responder group A and non-responder group B. The groups were compared and correlation between equiliberative nucleoside transporter expressions and liver fibrosis was evaluated. Data was analysed using SPSS 23. RESULTS: Of the 80 patients, 33(41.3%) were males and 47(58.8%) were females. The overall mean age was 37.46±10.61 years. In terms of response to treatment, there were 40(50%) in each of the two groups. Mean post-treatment duration was 15.38±30.09 weeks. Age was not significantly different with respect to gender (p>0.05), but the age pattern was significantly different between the two groups (p<0.001). Also, non-responders had significant post-treatment duration compared to the responders (p<0.001). Hypoxanthine-guanine phosphoribosyltransferase gene showed no significant difference between the groups (p=0.144). Equiliberative nucleoside transporter was significantly down-regulated in the non-responders (p<0.001) and showed correlation with the degree of liver fibrosis (p<0.034) compared to the responders. CONCLUSIONS: There was a significant association between equiliberative nucleoside transporters and liver fibrosis stage in hepatitis C virus non-responding patients.

Tumour infiltrating lymphocytes in colorectal cancers- correlation with tumour biology and clinical outcome: A cohort study.

OBJECTIVE: To evaluate the pattern of tumour infiltrating lymphocytes in colorectal cancers, and to correlate them with nuclear protein Ki67, vascular endothelial growth factor and clinical outcome. METHODS: The retrospective study was conducted at the Nuclear Institute of Medicine and Radiotherapy and the Liaquat University of Medical and Health Sciences, Jamshoro, Pakistan, and comprised data of colorectal cancer patients from January 1, 2008, to December 31, 2018. Whole tumour sections of colorectal cancer were used with haematoxylin and eosin staining, Histological type, grade and infiltrated lymphocytes within the tumour block were assessed. Ki67 and vascular endothelial growth factor were evaluated by immunohistochemistry, while the staining of these biomarkers was assessed by the percentage of cells stained. Data was analysed using SPSS 22. RESULTS: Of the 201 patients, 110(54.7%) were males and 91(45.3%) were females. Overall median age was 43 years (range 10-85 years). Majority of the tumours 132(65.7%) showed mild to moderate tumour infiltrating lymphocytes, 30(14.9%) had severe tumour infiltrating lymphocytes, while 39(19.4%) did not show any infiltrating lymphocytes. Tumour infiltrating lymphocytes did not show significant association with the histological grade (p>0.05), but high tumour infiltrating lymphocytes were associated with poor survival without being significantly associated with Ki67 pattern and vascular endothelial growth factor (p>0.05). CONCLUSIONS: Majority of colorectal cancer cases showed varying levels of lymphocyte infilteration, and tumour infiltrating lymphocytes were associated with poor survival, without having significant association with Ki67 pattern and vascular endothelial growth factor.

Evaluation of C-reactive protein and hematological parameters in smokeless tobacco users: A comparative cross-sectional study.

OBJECTIVE: To investigate the changes in levels of C-reactive protein (CRP) and hematological parameters among smokeless tobacco (SLT) users. METHODS: A comparative cross-sectional study was conducted at the community level in the coastal districts of Sindh province namely Badin, Thatta, and Sujawal from January 2017 to December 2019. The CRP and hematological parameters were evaluated by well-established methods among SLT and non-SLT users. RESULTS: There was a statistically significant difference between SLT users (mean CRP = 0.77) versus non-users (mean CRP = 0.18), p = <0.001. Among hematological parameters, white blood cells (SLT users median = 7.85 versus non-SLT users median = 8.50, p = 0.004), monocytes (SLT users median = 6.00 versus non-SLT users median = 6.00, p = 0.001) and erythrocyte sedimentation rate (SLT users median = 15.00 versus non-SLT users median = 10.00, p = 0.006) showed statistically significant difference. CONCLUSIONS: Significantly elevated CRP was observed in SLT users similarly hematological parameters also showed changes. WBCs, monocytes and ESR were significantly deranged among SLT users. Further studies looking into long term effects of these changes would be helpful.

LncRNA H19-mediated M2 polarization of macrophages promotes myofibroblast differentiation in pulmonary fibrosis induced by arsenic exposure.

Arsenic is a potent toxicant, and long-term exposure to inorganic arsenic causes lung damage. M2 macrophages play an important role in the pathogenesis of pulmonary fibrosis. However, the potential connections between arsenic and M2 macrophages in the development of pulmonary fibrosis are elusive. C57BL/6 mice were fed with drinking water containing 0, 10 and 20 ppm arsenite for 12 months. We have found that, in lung tissues of mice, arsenite, a biologically active form of arsenic, elevated H19, c-Myc, and Arg1; decreased let-7a; and caused pulmonary fibrosis. For THP-1 macrophages (THP-M) and bone-marrow-derived macrophages (BMDMs), 8 µM arsenite increased H19, c-Myc, and Arg1; decreased let-7a; and induced M2 polarization of macrophages, which caused secretion of the fibrogenic cytokine, TGF-ß1. Down-regulation of H19 or up-regulation of let-7a reversed the arsenite-induced M2 polarization of macrophages. Arsenite-treated THP-M and BMDMs co-cultured with MRC-5 cells or primary lung fibroblasts (PLFs) elevated levels of p-SMAD2/3, SMAD4, α-SMA, and collagen I in lung fibroblasts and resulted in the activation of lung fibroblasts. Knockout of H19 or up-regulation of let-7a in macrophages reversed the effects. The results indicated that H19 functioned as an miRNA sponge for let-7a, which was involved in arsenite-induced M2 polarization of macrophages and induced the myofibroblast differentiation phenotype by regulation of c-Myc. In the sera of arseniasis patients, levels of hydroxyproline and H19 were higher, and levels of let-7a were lower than levels in the controls. These observations elucidate a possible mechanism for arsenic exposure-induced pulmonary fibrosis.

Analysis of inflammatory markers in apparently healthy automobile vehicle drivers in response to exposure to traffic pollution fumes.

OBJECTIVE: This study aimed to evaluate pattern of markers of inflammation in apparently healthy drivers who exposed to traffic fumes. METHODS: This cross-sectional study was conducted from June 2016 to January 2017 at Liaquat University of Medical & Health Sciences (LUMHS), Jamshoro. It looked into the effects of traffic pollutants on markers of inflammation including CRP, Leukocytes count, IL-6, TNF-α, TNF-ß of healthy human volunteers. Eighty-seven, apparently healthy, non-smoking automobile vehicle drivers, having daily contact of traffic exhaust for at least six hours, aged between 18-40 years recruited for this study. Levels of traffic-generated pollutants P.M2.5, P.M10, NOx were recorded in different areas of Hyderabad City. RESULTS: P.M2.5 found to be positively correlated with markers of inflammation including IL-6 (rs = 0.99), TNF-α (rs = 0.41), CRP mg/dl (rs = 0.99) , neutrophils (rs = 0.29), lymphocytes (rs = 0.31), eosinophils (rs = 0.20), monocytes (rs = 0.42) and basophils (rs = 0.16). Positive correlation present among IL-6 (rs = 0.21), TNF-α (rs = 0.49) and CRP mg/dl (rs = 0.22) % (rs = -0.31), Leukocytes (rs = 0.14) neutrophils (rs = 0.31), lymphocytes (rs = 0.21), monocytes (rs = 0.50), basophils (rs = 0.17) with P.M10. NOx showed positive correlation with IL-6 (rs = 0.22), TNF-α (rs = 0.48), CRP (rs = 0.22), neutrophils (rs = 0.31), lymphocytes (rs = 0.13), basophils (rs = 0.17) and monocytes (rs = 0.48). CONCLUSION: Findings of our study suggest that almost all markers of inflammation are positively correlated with traffic pollutants and this condition might raise the risk of systemic diseases.

Effect of Body Mass Index on respiratory parameters: A cross-sectional analytical Study.

OBJECTIVE: To assess association of Body mass index (BMI) on respiratory parameters by performing spirometry in apparently healthy adults living in the district Jamshoro and Hyderabad, Sindh, Pakistan. METHODS: A cross sectional study was conducted at Department of Physiology, Liaquat University of Medical and Health Sciences Jamshoro, Pakistan from January to September 2015. A total of 180 underweight, normal, overweight and obese participants, aged between 18 to 40 years were included in the study. BMI was calculated by measuring weight and height by BMI scale (RGZ-160) in standing position. Pulmonary parameters were determined by spirometry on Power lab (AD instruments). Pulmonary parameters were compared between subjects in different categories of BMI. RESULTS: Mean age of participants was 21.83±5.88 years and the mean BMI was 25.10±6.55 kg/m2. The study results revealed that except for FVC, which was not statistically significant (p=0.45) all other respiratory parameters were significantly different (p≤0.05) in all BMI categories. Mean FEV1/FVC ratio (93.1 vs. 90.3, 86.4 and 86.6 respectively) was highest among underweight as compared to overweight, obese and normal weight individuals. The mean VT was 1.22 vs. 0.90, 1.01 and 0.84 respectively, IRV was 1.04 vs. 1.18, 1.23 and 1.20 respectively, IC was 2.26 vs. 2.08, 2.25 and 2.05 respectively, VC was 2.63 vs. 2.42, 2.54 and 2.54 respectively, TLC was 2.98 vs. 3.03 vs. 3.18 and 3.17 respectively among underweight, overweight, obese and normal weight participants. CONCLUSION: We found a significant association between body mass index and pulmonary function parameters. Obesity causes detrimental effects on respiratory system.

Liver Kinase B1-A Potential Therapeutic Target in Hormone-Sensitive Breast Cancer in Older Women.

Background: The role of liver kinase B1 (LKB1), a serine/threonine kinase, has been described in the development of PeutzJagher's syndrome, where a proportion (~45%) of patients have developed breast cancer in their lifetime. Cell line studies have linked LKB1 with oestrogen receptors (ER) and with the Adenosine monophosphate-activated protein kinase (AMPK) pathway for energy metabolism. However, limited studies have investigated protein expression of LKB1 in tumour tissues and its intracellular relationships. This study aimed to investigate the intracellular molecular relationships of LKB1 in older women with early operable primary breast cancer and its correlation with long-term clinical outcome. Methods: Between 1973 and 2010, a consecutive series of 1758 older (≥70 years) women with T0-2N0-1M0 breast carcinoma were managed in a dedicated facility. Of these, 813 patients underwent primary surgery, and 575 had good quality tumour samples available for tissue microarray construction. LKB1 was assessed in 407 cases by indirect immunohistochemistry (IHC). Tumours with 30% or more of cells with cytoplasmic LKB1 expression were considered positive. LKB1 expression was compared with tumour size, histological grade, axillary lymph node stage, ER, PgR, EGFR, HER2, HER3, HER4, BRCA1&2, p53, Ki67, Bcl2, Muc1, E-Cadherin, CD44, basal (CK5, CK5/6, CK14 and CK17) and luminal (CK7/8, CK18 and CK19) cytokeratins, MDM2 and MDM4, and correlated with long-term clinical outcome. Results: Positive LKB1 expression was seen in 318 (78.1%) patients, and was significantly associated with high tumour grade, high Ki67, over-expression of HER2, VEGF, HER4, BRCA2, MDM2 and negative expression of CD44 (p < 0.05). There was no significant correlation with tumour size, axillary lymph node status, ER, PgR, p53, basal or luminal cytokeratins, Bcl2, Muc1, EGFR, HER3, MDM4, E-cadherin and BRCA1. LKB1 did not show any significant influence on survival in the overall population; however, in those patients receiving adjuvant endocrine therapy for ER positive tumours, those with positive LKB1 had significantly better 5-year breast cancer specific survival when compared to those without such expression (93% versus 74%, p = 0.03). Conclusion: LKB1 expression has shown association with poor prognostic factors in older women with breast cancer. However, LKB1 expression appears to be associated with better survival outcome among those patients receiving adjuvant endocrine therapy. Further research is required to explore its potential role as a therapeutic target.

Correction: Oestrogen receptor negative early operable primary breast cancer in older women-Biological characteristics and long-term clinical outcome.

[This corrects the article DOI: 10.1371/journal.pone.0188528.].

Oestrogen receptor negative early operable primary breast cancer in older women-Biological characteristics and long-term clinical outcome.

BACKGROUND: Older women are at the greatest risk of breast cancer development and a considerable number present with comorbidities. Although the majority of breast cancers in this age group express oestrogen receptor (ER), which makes endocrine therapy (primary or adjuvant) feasible, given the huge size of the elderly population, there remains a significant number of patients, in absolute term, whose tumours do not express ER and their management is challenging. METHODS: Of a consecutive series of 1,758 older (≥70 years) women with early operable primary breast cancer managed in a dedicated service from 1973-2010, 252(14.3%) had ER-negative (histochemical (H) score ≤50) tumours. Their clinical outcome was retrospectively reviewed and tumour samples collected from diagnostic core biopsies were analysed for progesterone receptor (PgR), HER2 and Ki67 using immunohistochemistry. RESULTS: The commonest primary treatment was surgery (N = 194, 77%) followed by primary endocrine therapy (14.3%), primary radiotherapy (5.6%) and supportive treatment only (3.1%). Among the patients undergoing surgery, most of them had grade 3 (78.1%) and node-negative disease (62.2%). Some of them (21.1%) received postoperative radiotherapy. At a median follow-up of 37.5 months, 117 patients had died, out of which 48.6% were due to breast cancer. For those who underwent surgery, the regional and local recurrence rates were 2% and 1.1% per annum respectively. For those who received primary endocrine therapy, 38% progressed at 6 months, however all patients who had primary radiotherapy achieved clinical benefit at 6 months. Regardless of treatment given, the 5-year breast cancer specific and overall survival rates were 70% and 50% respectively. Biological analysis based on good quality needle core biopsy specimensfrom181 patients showed that 26.8% (N = 49), 16.9% (N = 31) and 70.7% (N = 70)expressed positivity for PgR, HER2 and Ki67 respectively. No correlation between these biomarkers and breast cancer specific survival was demonstrated. CONCLUSION: Oestrogen receptor negative early operable primary breast cancer in older women is associated with poor prognostic features in terms of biology and clinical outcome. Surgery appears to produce the best outcome as a primary treatment, however for those where neither surgery nor chemotherapy is appropriate, primary radiotherapy can be beneficial.