Effect of initial periodontal therapy on metallothionein levels in smokers and non-smokers with periodontitis.

The aim of this study was to determine the effect of non-surgical periodontal therapy (NSPT) on mRNA expression of metallothionein (MT) and its levels in serum, saliva and gingival crevicular fluid (GCF) of smokers (S) and non-smokers (NS) with periodontitis (P).A total of 100 participants were included: 48 periodontally healthy (PH) subjects (24 S [PH + S] and 24 NS [PH + NS]) and 52 patients with P (27 S [P + S] and 25 NS [P + NS]). Clinical parameters were recorded, and biofluids (serum, saliva and GCF) and gingival tissue samples were obtained at baseline in all groups and 3 months after NSPT in P groups. MT levels in biofluids were determined by ELISA. In gingival tissues, MT-mRNA expression was quantified using real-time PCR. mRNA expression of MT and its levels in biofluids were significantly higher in P + S compared to other groups, and the differences between P + NS and PH + S were non-significant. A significant decrease was observed for MT levels in biofluids, and MT-mRNA expression in periodontitis patients after NSPT. In conclusion, smoking and periodontitis are associated with higher MT expression which decreases after NSPT. MT as an oxidative stress biomarker and its therapeutic role in periodontitis should be investigated in future studies.Clinical trial registration: The study was prospectively registered at Clinical Trials Registry-India (ctri.nic.in) as CTRI/2018/08/015427 on August 23, 2018.

Expression analysis of MMP14: Key enzyme action in modulating visceral adipose tissue plasticity in patients with obesity.

Compromised adipose tissue plasticity is a hallmark finding of obesity orchestrated by the intricate interplay between various extracellular matrix components. Collagen6 (COL6) is well characterized in obese visceral adipose tissue (VAT), not much is known about MMP14 which is hypothesized to be the key player in matrix reorganization. Subjects with obesity (BMI ≥40; n = 50) aged 18-60 years undergoing bariatric surgery and their age-matched controls (BMI < 25; n = 30) were included. MMP14, Col6A3 and Tissue inhibitor of metalloproteinase 2 (TIMP2) mRNA expression was assessed in VAT and their serum levels along with endotrophin were estimated in both groups preoperatively and post-operatively in the obese group. The results were analysed statistically and correlated with anthropometric and glycaemic parameters, namely fasting glucose and insulin, HbA1c, HOMA-IR, HOMA-ß and QUICKI. Circulating levels as well as mRNA expression profiling revealed significant differences between the individuals with and without obesity (p < .05), more so in individuals with diabetes and obesity (p < .05). Follow-up serum analysis revealed significantly raised MMP14 (p < .001), with decreased Col6A3, endotrophin and TIMP2 levels (p < .01, p < .001 and p < .01, respectively). A rise in serum MMP14 protein, simultaneous with post-surgical weight loss and decreased serum levels of associated extracellular matrix (ECM) remodellers, suggests its crucial role in modulating obesity-associated ECM fibrosis and pliability of VAT.

An immediate post op and follow up assessment of circulating adipo-cytokines after bariatric surgery in morbid obesity.

BACKGROUND: Bariatric surgery has emerged as a promising treatment for improving adipose tissue dysfunction in obesity, but the mechanisms for such amelioration are still not known. This study comprehensively explores a panel of adipo-cytokines in individuals with obesity undergoing bariatric surgery, in conjunction with markers of insulin resistance, at three time points i.e., pre-op, immediate post-op and 6 months post-surgery. METHODS: It is a case-control prospective study among obese individuals undergoing bariatric surgery (BMI ≥35 kg/m2, n=30) and non-obese subjects (BMI <25 kg/m2, n=30), comparing the levels of serum adiponectin, resistin, C-Reactive Protein (CRP), Interleukin (IL)-6 and 8, Monocyte chemoattractant protein (MCP)-1 and Tumor necrosis factor (TNF)-α between them. The same were followed at immediate and 6-month post-op periods in the former group. The serum markers were correlated with the markers of Insulin resistance like HOMA-IR, HOMA-ß and QUICKI. RESULTS: A significant increase in adiponectin was seen after weight loss in obese group (17.54 ± 1.31 µg/mL at baseline vs 68.76 ± 1.84 µg/mL at 6- month post-surgery). CRP being an acute phase protein showed significant higher levels at immediate post-op period but declined even below its baseline at 6 months after surgery (33.34 ± 16.85 µg/mL at baseline vs 59.85 ± 23.12 µg/mL at immediate post-op vs 9.66 ± 1.84 µg/mL at 6 months post-operatively). Few inconsistencies were observed in the trajectories of IL-6 and TNF-α, while other pro-inflammatory markers indicated resolution after surgery. CONCLUSION: Bariatric surgery alleviated the systemic inflammation, correlating with improved insulin resistance in individuals with obesity.

Role of calcium ion channels and cytoskeletal proteins in Thorium-232 induced toxicity in normal human liver cells (WRL 68) and its validation in swiss mice.

Hepatic disorders reported in humans exposed to Thorium-232 (Th-232) rationalizes the present study investigating the toxicological response of normal human liver cells (WRL 68) and its validation in Swiss mice. Cell count analysis of WRL 68 cells-treated with Th-nitrate (1-200 µM) estimated IC50 of ∼24 µM (at 24 h) and 35 µM (at 48 h). Analysis of cell viability (trypan blue assay) showed the IC50 of ∼172 µM. Phase contrast bright-field microscopy revealed Th-induced morphological changes and cell-released microvesicle-like structures in extracellular space. Th-estimation by ICP-MS (Inductively-coupled plasma mass-spectrometry) showed uptake of Th by cells as a function of concentration and incubation time. Employing DTPA as a chelating agent in cell harvesting solution, cell-internalized/strongly-bound Th was estimated to be ∼42% of total incubated Th. Th-uptake studies in the presence of ion-channel specific inhibitors (e.g. nifedipine, thapsigargin) revealed the role of plasma membrane calcium channels and cytoplasmic calcium in modulating the Th-uptake. Transmission electron microscopy of Th-treated cells showed cell-derived extracellular vesicles, alterations in the shape and size of nucleus and mitochondria as well as cytoplasmic inclusions. The order of Th accumulation in various sub-cellular protein fractions was found to be as cytoskeleton (43%) > cytoplasmic (15%) > chromatin (7%) > nuclear (5%) & membrane (5%). Immunofluorescence analysis of WRL 68 cells showed that Th significantly altered the expression of cytoskeleton proteins (F-actin and keratin), which was further validated in liver tissues of Swiss mice administered with Th-232. Findings herein highlight the role of calcium channels and cytoskeleton in Th-induced toxicity. Keywords: Thorium toxicity; Liver cells; Calcium channels; Sub-cellular targets, Cytoskeleton; Swiss Mice.

Metallothionein levels in gingival crevicular fluid, saliva, and serum of smokers and non-smokers with chronic periodontitis.

BACKGROUND: Metallothionein (MT), a cysteine rich protein is involved as a radical scavenger in several pathological conditions associated with oxidative stress; however, its role in periodontal disease still remains elusive. The aim of this cross-sectional study is to determine the serum, saliva and gingival crevicular fluid (GCF) levels of MT in smokers (S) and non-smokers (NS) with chronic periodontitis (CP), and compare them with those of periodontally healthy (PH) individuals. METHODS: A total of 85 participants were enrolled: 45 patients with CP (23 S [CP+S] and 22 NS [CP+NS]) and 40 PH individuals (20 S [PH+S] and 20 NS [PH+NS]). In all the study participants, clinical periodontal parameters (plaque index, gingival index, sulcus bleeding index, probing depth, and clinical attachment level) were recorded and samples of serum, saliva and GCF were collected. Enzyme-linked immunosorbent assay was used to determine the levels of MT in the samples. RESULTS: All periodontal clinical parameters were significantly higher in the CP groups as compared to PH groups (P < 0.05). MT levels in CP+S group were significantly raised in comparison to other three groups. There was no statistically significant difference in MT levels among CP+NS and PH+S groups (P > 0.05); however, relatively higher levels were observed in GCF and saliva in CP+NS group. When all the study groups were observed together, MT levels were positively correlated with clinical parameters. CONCLUSIONS: Results of present study suggest that smoking and CP can induce the synthesis of MT owing to increased oxidative stress and heavy metals intoxication. Further longitudinal studies with large sample size and an interventional arm are needed to substantiate the role of MT as a potential biomarker in periodontitis.

Serum Adipocytokines Levels and Their Association with Insulin Sensitivity in Morbidly Obese Individuals Undergoing Bariatric Surgery.

BACKGROUND: Obese adipose tissue secretes a variety of adipocytokines that act as metabolic regulators with complex mechanisms. Our objective was to compare serum concentration of a panel of adipocytokines between obese and non-obese individuals and identify any distinct patterns correlating with insulin sensitivity in obesity. METHODS: We designed a cross-sectional study among obese (body mass index [BMI] ≥30 kg/m2, n=62) and non-obese (BMI <25 kg/m2, n=32) individuals to compare circulating levels of the adipokines, such as adiponectin and resistin in conjunction with the measurement of the levels of inflammatory cytokines including C-reactive protein (CRP), interleukin (IL)-6, IL-8, monocyte chemoattractant protein (MCP)-1, and tumor necrosis factor (TNF)-α using Luminex multiplex immunoassay with drop array technology. Correlations between circulating adipocytokine levels and those of multiple well-established markers of insulin resistance including homeostatic model assessment of insulin resistance (HOMA-IR), homeostatic model assessment of ß-cell function (HOMA-ß) and quantitative insulin sensitivity check index were also established. RESULTS: CRP, IL-8, MCP-1, and TNF-α levels were higher in obese than non-obese individuals; the CRP and IL-8 differences were statistically significant. CRP correlated significantly with markers of insulin resistance (fasting plasma insulin, HOMA-IR, and QUICKI), and adiponectin correlated with HOMA-ß in obese individuals. We divided the group of obese individuals on the basis of HOMA-IR levels into insulin-resistant (IR; HOMA-IR ≥2.5) and insulin-sensitive (IS; HOMA-IR <2.5) groups; and 43 out of 62 participants were IR despite comparable BMIs. An overall proinflammatory profile was compared between IR and IS obese, though the values were higher in IR obese but the difference was not significant. CONCLUSION: Obesity is associated with a general inflammatory milieu and a crosstalk between adipocytokines and insulin resistance is complex as well as multifactorial.

SARS-CoV-2-host dynamics: Increased risk of adverse outcomes of COVID-19 in obesity.

BACKGROUND AND AIM: The pandemic of COVID-19 has put forward the public health system across countries to prepare themselves for the unprecedented outbreak of the present time. Recognition of the associated risks of morbidity and mortality becomes not only imperative but also fundamental to determine the prevention strategies as well as targeting the high-risk populations for appropriate therapies. METHODS: We reviewed, collated and analysed the online database i.e. Pubmed, Google scholar, Researchgate to highlight the demographic and mechanistic link between obesity and associated risks of severity in COVID-19. RESULTS: We observed a changing dynamic in the reporting from the time of initial pandemic in China to currently reported research. While, initially body mass index (BMI) did not find a mention in the data, it is now clearly emerging that obesity is one of the profound risk factors for complications of COVID-19. CONCLUSION: Our review will help clinicians and health policy makers in considering the importance of obesity in making the prevention and therapeutic strategies of COVID-19. An extra attention and precaution for patients with obesity in COVID-19 pandemic is recommended.

Estimation and in-situ detection of thorium in human liver cell culture by arsenazo-III based colorimetric assay.

To understand the biological effects of Thorium-232 (Th) in human cells and animal models as well as to assess mitigation strategies for its detoxification, there is a need to develop a sensitive, specific, high-throughput and easily-implementable assay for detection and estimation of Th in biological samples. Here, we have optimized arsenazo-III dye based colorimetric assay to detect Th in biological samples. The concentration of arsenazo-III (i.e. 50 µM) was optimized, which can reliably estimate Th in the concentration range of 2.5 to 40 µM. The optimized assay can specifically detect Th without interference from other metal ions (La, Ce, U, Fe, Ca, Cu, Zn and Mn). A significant correlation (R2 = 0.999) was found between arsenazo-III-based detection of Th and total reflection X-ray fluorescence. The conditions of present assay successfully estimated Th in cell culture medium, cell harvesting (trypsin-EDTA) solution and cell lysate obtained from human liver cell culture. Moreover, for the first time, we detected Th in-situ in adherent liver cells in culture after staining with arsenazo-III. This study confirms that Th can be specifically determined in biological samples using arsenazo-III with the sensitivity, which is relevant to thorium toxicity research.

DNA structural basis for fragility at peak III of BCL2 major breakpoint region associated with t(14;18) translocation.

Maintaining genome integrity is crucial for normal cellular functions. DNA double-strand breaks (DSBs), when unrepaired, can potentiate chromosomal translocations. t(14;18) translocation involving BCL2 gene on chromosome 18 and IgH loci at chromosome 14, could lead to follicular lymphoma. Molecular basis for fragility of translocation breakpoint regions is an active area of investigation. Previously, formation of non-B DNA structures like G-quadruplex, triplex, B/A transition were investigated at peak I of BCL2 major breakpoint region (MBR); however, it is less understood at peak III. In vitro gel shift assays show faster mobility for MBR peak III sequences, unlike controls. CD studies of peak III sequences reveal a spectral pattern different from B-DNA. Although complementary C-rich stretches exhibit single-strandedness, corresponding guanine-rich sequences do not show DMS protection, ruling out G-quadruplex and triplex DNA. Extrachromosomal assay indicates that peak III halts transcription, unlike its mutated version. Taken together, multiple lines of evidence suggest formation of potential cruciform DNA structure at MBR peak III, which was also supported by in silico studies. Thus, our study reveals formation of non-B DNA structure which could be a basis for fragility at BCL2 breakpoint regions, eventually leading to chromosomal translocations.

A Study of the Extended Lipid Profile including Oxidized LDL, Small Dense LDL, Lipoprotein (a) and Apolipoproteins in the Assessment of Cardiovascular Risk in Hypothyroid Patients.

INTRODUCTION: Hypothyroidism is one of the most common metabolic disorders associated with dyslipidemia which poses a higher risk of Coronary Artery Disease (CAD) in such patients. Biochemical markers which can pick up the risk promptly are becoming imperative now-a-days and thus the assessment beyond the conventional lipid profile is the need of the hour. AIMS: To assess the association of non-conventional lipid parameters like small dense Low Density Lipoprotein (sd LDL), oxidized Low Density Lipoprotein (ox LDL), Apolipoprotein A (Apo A1), Apolipoprotein B (Apo B) and Lipoprotein (a) {Lp(a)} in hypothyroid patients and compare their values with the conventional lipid parameters such as Total Cholesterol (TC), Triglyceride (TG), Low-Density Lipoprotein Cholesterol (LDL-C) and High-Density Lipoprotein Cholesterol (HDL-C). MATERIALS AND METHODS: One hundred and thirty clinically proven patients of hypothyroidism aged 20-60 years and equal number of age and gender matched healthy individuals were included in this case control study. Serum sd LDL, ox LDL, Apo A1, Apo B, Lp (a), lipid profile, Thyroid Stimulating Hormone (TSH), Free Triiodothyronine (FT3) and Free Tetraiodothyronine (FT4) levels were measured in both the groups. The data was recorded and analysed on SPSS system. The results of cases and controls were compared by student t-test and one-way ANOVA. All the parameters were correlated with TSH by Pearson's correlation. RESULTS: We found significantly high levels of sd LDL, ox LDL, Apo B, Lp (a), TC, TG, LDL-C in cases as compared to the controls. Ox LDL has shown maximum correlation with serum TSH (p<0.0001, r=0.801) followed by sd LDL (p<0.0001, r=0.792), Apo B (p<0.001, r=0.783) and LDL-C (p<0.001, r=0.741). Moreover, ox LDL and sd LDL were found to be increased in normolipidemic hypothyroid patients thereby giving a strong supportive evidence that estimation of these parameters can become fundamental in prompt identification of the high risk patients of CAD in hypothyroid population. CONCLUSION: Non-conventional lipid parameters appear to be better markers for the assessment of cardiovascular risk in hypothyroidism and might help in the designing of the effective treatment protocols and areas of intervention by the clinicians as well as researchers.

The evaluation of serum amylase in the patients of type 2 diabetes mellitus, with a possible correlation with the pancreatic functions.

BACKGROUND: Diabetes mellitus is a chronic metabolic disorder which is associated with hyperglycaemia. It is caused by a derangement in the secretion or function of the endocrinal portion of the pancreas. There is a close anatomical and functional relationship between its exocrine and endocrine portions. To address this issue, the current study was designed to evaluate the blood glucose and the amylase levels of diabetic patients as representatives of the two portions of the pancreas respectively. AIMS AND OBJECTIVES: The aim of the present study was to determine the blood glucose, serum amylase and the serum lipid profile in known cases of type 2 Diabetes Mellitus and to compare and correlate these parameters with those of age and sex matched healthy controls. MATERIAL AND METHODS: One hundred ten patients of type 2 Diabetes mellitus, who were already diagnosed and were taking treatment, were included in this study. 30 age and sex matched healthy individuals were recruited as the control group in our study. Fasting venous blood samples were collected from the patients as well as the controls and they were analysed by using an automated analyser for blood glucose, serum amylase and the lipid profile (serum triglycerides, total cholesterol, high density lipoproteins and low density lipoproteins). The results were analyzed statistically by using the Student's "t" test and correlation coefficients. RESULTS: Significantly low serum amylase levels were found in the diabetic patients as compared to those in the healthy controls (p value <0.001). Also, the levels of fasting serum total cholesterol, triglycerides and the low density lipoproteins were significantly higher in the patients as compared to those in the controls, with p values of <0.05, <0.001 and <0.001 respectively. The HDL (high density lipoprotein) level was found to be lower in the diabetic patients (p value <0.001). CONCLUSIONS: From our study, it was concluded that in type 2 Diabetes mellitus, wherever the blood glucose level was higher, the serum amylase activity was found to be significantly lower. This reflected the derangement in the endocrine-exocrine axis of the pancreas, as a disease which affected any portion of an organ would affect the adjoining area of that organ functionally. This fact must be kept in mind while the patients are treated.

Association of insulin like growth factor-1 (IGF-1) and thyroid hormones in patients of acute leukemia.

BACKGROUND: For many years, several studies have demonstrated a relationship between insulin like growth factor-1 (IGF-1), thyroid hormones, and various malignancies. IGF-1 plays an important role in tumor proliferation in various malignancies. The relationship between IGF-1 and thyroid hormones is complex and not fully understood. Therefore we planned to evaluate the level of IGF-1 and thyroid hormones in patients of acute leukemia. METHODS: The present study included 25 patients with acute leukemia (Acute Myeloid Leukemia, n = 16; Acute Lymphoid Leukemia n = 9, mean age 28.16 years). 25 age and gender matched healthy individuals were taken as control (mean age 27.17 years). In all the subjects, serum IGF-1 was measured by enzyme linked immunosorbent assay (ELISA), serum total triiodothyronine (T3), thyroxine (T4) by radioimmunoassay (RIA), thyroid stimulating hormone (TSH) by immunoradiometric assay (IRMA), and free T3 (FT3) and free T4 (FT4) by chemilluminiscence. These tests were done before starting of chemotherapy and either 6 to 8 weeks after chemotherapy or at the time of remission, whichever was earlier. RESULTS: At the time of diagnosis, patients with acute leukemia showed a significantly increased level of IGF-1 as compared to controls (198.32 +/- 67.55 vs 160.64 +/- 45.96; p < 0.01). After 6 to 8 weeks of chemotherapy, patients with acute leukemia showed a significant decrease in the level of IGF-1 compared to the baseline values (198.32 +/- 67.55 vs 155.6 +/- 45.96; p < 0.01). Though FT3, FT4, total T3, and total T4 values in these patients were within the normal range, these values were still significantly higher compared to controls. TSH levels were significantly lower in patients at the time of presentation and the levels increased after chemotherapy. CONCLUSIONS: The estimation of IGF-1 and thyroid hormones may be helpful in assessing the disease activity and predicting the response of chemotherapy.