An association of VDR gene polymorphism in hypovitaminosis D mediated secondary hyperparathyroidism in adolescent girls; a tertiary hospital study in central India.

Vitamin D is required to maintain normal serum calcium and phosphate levels that help normal bone mineralization, nerve conduction, muscle contraction, immune function, cell proliferation, and differentiation. Interventions including vitamin D supplementation may not improve vitamin D deficiency, as various complex genomic actions could contribute to vitamin D deficiency in the Indian population. Thus, we assessed hypovitaminosis D's relationship with vitamin D receptor (VDR) gene polymorphism and evaluated parathyroid hormone (PTH) levels in seemingly healthy adolescent school-going girls. We included 100 school-going girls (aged 12-17 years) studying in four schools of different socio-economic strata of Bhopal, India. The selected participants were divided into four groups based on the school in which they were studying. Blood samples were tested for serum calcium, phosphorus, PTH, ALP, vitamin D 25(OH) D, and albumin levels.VDR polymorphism was detected through the PCR-RFLP. Data were analyzed using the chi-square test, ANOVA, and linear regression. The difference in the age, calcium, ALP, and vitamin D values between the four groups were significant (P < 0.05), whereas high PTH levels (80%) were found. A higher prevalence of homozygous polymorphic allele demonstrates a molecular signature for severe secondary hyperparathyroidism. Hypovitaminosis D ranged from 84.9% to 100%, and a high prevalence of VDR polymorphism was observed. Attention must be paid to the health of this age group of school-going girls as hypovitaminosis D and associated VDR gene polymorphism could be the reason for secondary hyperparathyroidism (SHPT), showing changes in bone mineral density in these adolescent girls to ensure their future health.

Metabolic syndrome in non-obese patients with OSA: learning points of a cross-sectional study from a tertiary care hospital in Central India.

STUDY OBJECTIVES: Obesity is often considered mandatory for the diagnosis of Metabolic Syndrome (MS). Data on the prevalence of MS in non-obese patients with Obstructive Sleep Apnea (OSA) is scarce. This study was aimed to determine the prevalence of MS in non-obese patients with OSA. METHODOLOGY: All consecutively diagnosed patients with OSA between October 2018 and November 2019 were screened for metabolic syndrome. Patients with OSA and BMI < 25 kg/m2 (NOOSA) vs BMI > 25 kg/m2 (obese OSA) were compared. Lean waist NOOSA was defined as BMI < 25 kg/m2 and WC < 80 cm (32 in.) for women or < 90 cm (36 in.) for men. RESULTS: During the study period, 502 patients were diagnosed with OSA. MS was observed in 35% of patients with NOOSA compared to obese patients with OSA (79%). In the NOOSA group, hypertension, impaired fasting glucose, diabetes mellitus and dyslipidemia were observed in 65, 48, 14 and 61% respectively and all of these parameters were significantly more common in the obese group (p < 0.001). Parameters of OSA severity (apnea-hypopnea index or AHI, time spent below 90% saturated or T90, and nadir oxygen) were significantly more severe in the obese group with OSA. Approximately 83% of patients in the NOOSA group had at least two metabolic risk factors, compared to the obese OSA group, in which 95% had two or more metabolic risk factors. Sixty-four percent of patients with NOOSA with lean waist had at least two metabolic risk factors. At BMI cut-offs of < 25, < 27 and < 30 kg/m2; 35, 46 and 57% of patients with OSA respectively had metabolic syndrome. CONCLUSION: Metabolic syndrome was observed in approximately one in three patients with OSA and BMI < 25 kg/m2. Approximately two of every three lean waist non-obese patients with OSA had at least two markers of metabolic syndrome. The role of OSA in the development of metabolic syndrome in non-obese individuals needs further exploration.

Women with OSA have higher chances of having metabolic syndrome than men: effect of gender on syndrome Z in cross sectional study.

STUDY OBJECTIVES: This study was done to find out prevalence of Metabolic syndrome (MS) in patients with Obstructive Sleep Apnea (OSA) and whether there is any difference in prevalence of syndrome Z in male and female. METHODOLOGY: All consecutive diagnosed patients with OSA between June 2015 and Oct 2019 were screened for metabolic syndrome and factors associated with metabolic syndrome in OSA were analyzed. RESULTS: During study period, 502 patients (357 males; 145 females) were diagnosed with OSA. Mean age was 51.88 ± 12.18 years (females and males:55.91 ± 9.74 and 50.24 ± 12.70 years, respectively). Mean BMI was 31.60 ± 11.09 kg/m2 (female: 35.29 ± 7.19 and male: 30.1 ± 12.0 kg/m2) (p < 0.001). Mean AHI was 62.67 ± 35.22. Mild, moderate and severe category of OSA constituted 7.3%, 15.3% and 77.4% respectively. MS was found in 72.7% (365 out of 502) individuals with OSA. MS was found in 75.8%, 68.4 and 48.7% in severe, moderate and mild OSA patients respectively (p < 0.001). Females OSA patients had significantly high percentage (88.27%) of metabolic syndrome compared to males OSA patients (66.38%) {p < 0.001}. Female patients with SZ had higher metabolic score (p = 0.019) and were older (p < 0.001). CONCLUSION: Metabolic syndrome is highly prevalent in OSA population (72.7%) and is much more common in female OSA patients (88%) than males OSA (68%). All OSA patients should be screened for MS so that early intervention can be done in these patients so as to prevent cardiovascular complications.

Abnormal Ambulatory Blood Pressure and Early Renal Dysfunction in Sickle Cell Disease.

OBJECTIVES: There is sparsity of studies evaluating blood pressure in children with sickle cell disease (SCD), which have shown inconsistent results. Few of the studies have documented lower office blood pressure (BP) in SCD patients, whereas, others have shown presence of masked hypertension and abnormal ambulatory blood BP monitoring (ABPM). Thus, the present study was conducted to examine 24 h ABPM parameters and renal dysfunction in children with SCD and compare them with healthy controls. METHODS: A cross-sectional study was conducted on 56 children (30 children having SCD and 26 controls). ABPM and evaluation of renal functions including serum creatinine, serum urea, urinary creatinine, urinary protein and specific gravity was performed. RESULTS: Spot urinary protein to creatinine ratio was found to be higher in patients with SCD (63.3%) as compared to controls (p < 0.001). Proteinuria was observed in 1/4th of the SCD patients less than ten years of age. Masked hypertension was present in 2 (6.6%) patients, ambulatory hypertension in 4 (13.3%), ambulatory pre-hypertension in 1 (3.3%) and abnormal dipping in 60%. A statistically significant correlation of BMI for age Z-score and standard deviation score (SDS/Z) of 24 h systolic BP (r = 0.56, p = 0.002); estimated glomerular filtration rate (eGFR) with 24 h diastolic BP SDS (r = -0.52; p = 0.038) and age with e GFR (r = 0.54; p = 0.025) was found in the present study. CONCLUSIONS: The present study corroborates that ABPM abnormalities (ambulatory hypertension, non-dipping pattern, ambulatory prehypertension) and early onset proteinuria are significant findings in patients with SCD. This underscores the importance of regular screening for proteinuria and ABPM in routine care, for early detection and prevention of progressive renal damage in SCD.

Evaluation of MTBDRsl for detecting resistance in Mycobacterium tuberculosis to second-line drugs.

SETTING: Patients with presumed multidrug-resistant tuberculosis (MDR-TB) and undergoing MDR-TB treatment from Rajasthan, India.OBJECTIVE: To compare the GenoType® MTBDRsl v.1.0 (MTBDRsl) assay capacity to detect resistance to ofloxacin, amikacin, capreomycin, kanamycin and ethambutol in Mycobacterium tuberculosis with phenotypic drug susceptibility testing (DST) using MGIT™960™ in sputum samples and isolates.DESIGN: Fifty-three smear-positive sputum samples were tested directly by MTBDRsl and 205 MDR-TB isolates were processed using MTBDRsl and DST for five drugs on MGIT960. DNA sequencing was performed in isolates with discordance in the results between the two methods for the gyrA, gyrB and rrs genes.RESULT: Sensitivity and specificity of MTBDRsl was found to be respectively 93.1% and 100% for fluoroquinoline, respectively 75-78% and 100% for aminoglycosides/cyclopeptides, respectively 70% and 92% for ethambutol and respectively 92.3% and 100% for extensively drug-resistant (XDR) TB detection. On sequencing eight discordant isolates for quinolones, mutations were seen in 12.5% of the gyrB gene and among 20 discordant isolates for aminoglycosides/cyclopeptides in the rrs gene in 15% isolates. The turnaround time was 2 days for MTBDRsl vs. 10 days for MGIT960.CONCLUSIONS: MTBDRsl can be used as an initial rapid test for detecting XDR-TB, resistance to quinolones and aminogycosides/cyclopeptides in smear-positive sputum samples.

Occupational health hazards of trichloroethylene among workers in relation to altered mRNA expression of cell cycle regulating genes (p53, p21, bax and bcl-2) and PPARA.

Trichloroethylene (TCE) is widely used as a metal degreaser in industrial processes. The present study reports on the effects of TCE exposure on workers employed in the lock industries. To ensure exposure of the workers to TCE, its toxic metabolites, trichloroacetic acid (TCA), dichloroacetic acid (DCA) and trichloroethanol (TCEOH) were detected in the plasma of the subjects through solid phase microextraction-gas chromatography-electron capture detection. TCA, DCA and TCEOH were detected in the range of 0.004-2.494 µg/mL, 0.01-3.612 µg/mL and 0.002-0.617 µg/mL, respectively. Quantitative reverse transcription polymerase chain reaction analysis revealed up-regulated expression of p53 (2.4-fold; p < 0.05), p21 (2-fold; p < 0.01), bax (2.9-fold; p < 0.01) mRNAs and down-regulated expression of bcl-2 (67%; p < 0.05) mRNAs, indicating DNA damaging potential of these metabolites. No effects were observed on the levels of p16 and c-myc mRNAs. Further, as TCA and DCA, the ligand of peroxisome proliferator activated receptor alpha (PPARA), are involved in the process of hepatocarcinogenesis in rodents, we examined expression of PPARA mRNA and let-7c miRNA in the workers. No statistically significant differences in expression of PPARA mRNA and let-7c miRNA in patients were observed as compared to values in controls. Dehydroepiandosterone sulfate (DHEAS) is a reported endogenous ligand of PPARA so its competitive role was also studied. We observed decreased levels of DHEAS hormone in the subjects. Hence, its involvement in mediation of the observed changes in the levels of various mRNAs analyzed in this study appears unlikely.

In vitro cytogenetic assessment of trichloroacetic acid in human peripheral blood lymphocytes.

Trichloroacetic acid (TCA), a common water disinfection byproduct and a persistent metabolite of trichloroethylene (TCE), has been examined for its genotoxic potential in human lymphocytes. Chromosomal aberration (CA) and cytokinesis-block micronucleus (CBMN) assay were employed to assess the toxicity of TCA. Lymphocytes obtained from three healthy donors were exposed to 25, 50, and 100 µg/ml concentration of TCA separately. TCA exposure resulted in chromosomal anomalies and the formation of micronuclei in lymphocytes. Chromosome analysis revealed the dose-dependent and significant induction of CA. Chromatid break/chromosome break, fragments, and chromatid exchanges were commonly observed. Exposure of higher concentration (50 and 100 µg/ml) significantly inhibited mitotic index. Data obtained with CBMN assay indicated that the induction of micronucleus (MN) formation was greater than that of CA. At 25 µg/ml, TCA induced significant frequencies of MN as compared to control cells. Significant induction of MN at the lowest concentration indicates TCA may also interact with mitotic spindles. Lower percentage of CA and MN at 100 µg/ml as compared to 50 µg/ml indicates occurrence of severe cytotoxicity on exposure of 100 µg/ml TCA in lymphocytes. Collectively, results of both cytogenetic assays indicate that exposure of TCA can induce significant genotoxic and cytotoxic effects.

Myeloma cast nephropathy: a rare cause of primary renal allograft dysfunction.

We describe a rare case of primary renal allograft dysfunction due to myeloma cast nephropathy in a patient with no prior history of multiple myeloma preceding her transplantation. A 72-year-old woman on hemodialysis for 4 years prior to transplantation due to presumed hypertensive nephrosclerosis developed immediate graft dysfunction posttransplantation. The graft biopsy specimens were consistent with myeloma cast nephropathy for which she was treated with plasmapheresis and chemotherapy using bortezomib and dexamethasone. She achieved partial hematologic remission and recovered excellent graft function by 3 months posttransplantation. This is the first report of successful recovery of renal allograft function after development of cast nephropathy.

Antibody response to purified chick embryo cell vaccine in equines for production of equine rabies immune globulin.

Rabies is endemic in India. The post exposure treatment of class three bite cases, as recommended by the World Health Organization, must involve the use of rabies immunoglobulin as soon as possible and up to the seventh day of start of anti rabies vaccination. The annual requirement in India as projected by the Ministry of Health, Government of India is about 1500 liters of purified anti rabies serum (ERIG). Central Research Institute (CRI), Kasauli, being the sole producer of ERIG in India, an effort was made to increase the production of ERIG by the use of tissue culture vaccine (Human) for primary immunization of equines. It also involved changing the vaccine from horse brain suspension to tissue culture vaccine by eliminating horse sacrifice for antigen preparation. A better immune response was obtained.

Application of the PCR technique for a rapid, specific and sensitive detection of Salmonella spp. in foods.

We report here the use of a PCR based assay modified by us for the detection of Salmonella spp. in foods, based on amplification of a 236 bp Salmonella specific hin/H2 region [Way et al. (1993) Applied and Environmental Microbiology 59, 1473-1479], using Ampli Taq Gold polymerase. Using this assay we were able to detect all the Salmonella serovars tested. The limit of detection was 1 fg of purified target DNA or N x 10(0) (1-3 cells) cfu ml(-1) of pure bacterial culture. This assay could detect N x 10(0) cfu Salmonella cells g(-1) of the food sample unambiguously in presence of endogenous microflora following 6 h enrichment, thus requires a duration of approximately 10 h for the full processing from DNA template preparation, PCR and visualization of DNA product on agarose gel. The main advantage of this PCR detection method is its sensitivity, and specificity. We also tried to adopt DNA template isolation method simply by boiling the bacterial cells thereby reducing the possibility of contamination, cutting the processing time and cost considerably. This can be an added advantage for the use of this system in simple lab setups.

Absolute Doubly Differential Cross Section for Bremsstrahlung Spectra Produced in 7.0 keV e

The first experimental data are presented for the absolute doubly differential cross section (DDCS) for non-characteristics (bremsstrahlung) X-ray spectra produced by 7.0 keV electron bombardment of (semi-thick) targets of silver and gold at a photon detection angle of 90 degree. The bremsstrahlung spectra are corrected for detector's efficiency as well as for target effects; namely, electron energy loss, backscattering and photon-attenuation in the target. The DDCS values so obtained are compared with the predictions of a thin target bremsstrahlung theory. The agreement between experiment and theory for DDCS both in magnitude and shape is found to be satisfactory within the systematic experimental uncertainty of about 27%. penetrating electrons into the thick targets and the depth distribution of photons generated inside the targets are expected to be also present in the interaction. However, the contributions of these effects to the DDCS have not been taken into consideration. Various possible reasons for the existing discrepancy between experiment and theory are pointed out.

Trichloroethylene toxicity in mice: a biochemical, hematological and pathological assessment.

Oral administration of trichloroethylene (TCE; 0, 500, 1000 and 2000 mg/kg/day) to male mice once daily, 5 days a week for a period of 28 days, caused a significant increase in liver weight, degeneration/necrosis of hepatocytes and characteristics proliferation of endothelial cells of hepatic sinusoids. Increase in kidney weight, glomerular nephrosis, degeneration/desquamation of tubular epithelium and characteristic amyloid deposition in glomeruli were observed only in the group of mice treated with 2000 mg/kg TCE. These changes occurred concurrently with a significant increase in total protein and free sulphydryl contents, elevated activities of acid phosphatase and catalase and decreased activity of delta-aminolevulinic acid dehydratase (delta-ALAD) indicating the sensitivity of liver and kidney as target tissues in TCE-toxicity. Hematological studies showed a significant increase in RBC counts and a reduction in WBC counts without any statistically significant change in the hemoglobin, urea nitrogen, creatinine and uric acid levels in the blood of TCE-exposed mice. A dose-related increase in cell density and acid phosphatase activity with a parallel significant decrease in the activity of delta-ALAD were observed in the bone marrow, which appear to be responsible for hematological alterations in TCE-exposed mice. The results suggest that early metabolic, pathological and hematological perturbations following a short-term exposure of TCE in mice, can provide the basis for its documented potential for chronic effects like blood dyscrasia and cancer.

Influence of trichloroethylene treatment on phosphoinositides in rat brain.

The effect of oral administration of trichloroethylene, a neurotoxic solvent, on the levels of phosphoinositides in rat brain was studied. Two hours after administration of a single dose of trichloroethylene (1000 mg/kg body wt.), the levels of phosphatidylinositol (PI) and phosphatidylinositol 4,5-biphosphate (PIP2) were reduced by 24 and 17%, respectively, without any significant change in that of phosphatidylinositol-4-phosphate (PIP). Twenty hours after treatment, the levels of PI and PIP2 were increased by 22 and 38%, respectively. Repeated administration of the same dose of trichloroethylene for 1 year markedly reduced the levels of PI (52%), PIP (23%) and PIP2 (45%). These results for the first time suggest the involvement of a phosphoinositide messenger system in trichloroethylene neurotoxicity.

Hepatotoxic effects elicited by n-hexane or n-heptane.

Hepatotoxic effects of n-hexane and n-heptane administered i.p. (1 ml/kg body wt) were studied in albino rats after 1, 2, 7 and 45 days of treatment. Hepatic protein content decreased with n-heptane and total sulphydryl content showed a significant decrease in the rats exposed to either solvent. A significant increase in lipid peroxidation was observed after 24 h and 48 h exposure to n-hexane or n-heptane. A marked decrease in drug metabolizing activity and an increase in pentabarbitone sleeping time was also observed. Hepatic glucose-6-phosphatase, a microsomal marker enzyme, showed a significant decrease.

Toxicity of n-hexane and 2,5-hexanediol to the haemopoietic system: a preliminary report.

The effect of n-hexane and its metabolite, 2,5-hexanediol, on the haemopoietic system has been investigated in the rat. 59Fe uptake was decreased in the bone marrow of n-hexane-treated animals. Spleen and bone marrow were the target sites for the metabolite 2,5-hexanediol, whereas n-hexane as such does not affect these organs.

Peroxisome proliferation and lipid peroxidation in rat liver.

Male F344 rats were fed a diet containing the peroxisome proliferators 2-[4-(2,2-dichlorocyclopropyl)phenoxy]-2-methylpropionic acid [ciprofibrate (0.025%)] or [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid [Wy-14643 (0.1%)] for up to 14 months to determine whether hepatic peroxisome proliferation caused by these agents results in the induction of membrane lipid peroxidation in the liver. Peroxidative damage of membrane lipids from whole liver, postnuclear, heavy-particle, microsomal, and nuclear membranes was evaluated by determining the extent of formation of conjugated dienes (ultraviolet absorption, 233 nm). Increased generation of diene conjugates was noted in whole-liver, postnuclear, and heavy-particle membrane lipids of rats fed peroxisome proliferators for 6 months or longer when compared to controls. An additional, more intense absorption profile in the ultraviolet absorption range of approximately 276 nm was noted in the membrane lipids derived from whole liver, postnuclear, and heavy particle pellets, but not in the nuclear and microsomal membrane lipids of livers with peroxisome proliferation. Although the exact chemical nature of this delta 276 nm peak is not clear, it is attributed to the formation of ketone dienes and/or conjugated trienes. The excess lipid peroxidation correlates with the previous observation of accumulation of abundant quantities of lipofuscin in hepatocytes of rats chronically exposed to peroxisome proliferators. The generation of conjugated dienes and ketone dienes and/or trienes together with increased levels of H2O2 generation by peroxisomal enzymes, and decreased levels of hepatic glutathione peroxidase, glutathione reductase, and glutathione-S-transferases, enzymes responsible for the defense against H2O2 damage, suggest the occurrence of membrane lipid peroxidation and oxidative stress in livers of rats treated with carcinogenic peroxisome proliferators.

Transcription regulation of peroxisomal fatty acyl-CoA oxidase and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase in rat liver by peroxisome proliferators.

The structurally diverse peroxisome proliferators ciprofibrate, clofibrate, and bis(2-ethylhexyl) phthalate [(EtHx)2 greater than Pht] increase the activities of hepatic catalase and peroxisomal fatty acid beta-oxidation enzymes in conjunction with profound proliferation of peroxisomes in hepatocytes. In order to delineate the level at which these enzymes are induced in the liver, the transcriptional activity of specific genes for fatty acyl-CoA oxidase (FAOxase) and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme (PBE), the first two enzymes of the peroxisomal beta-oxidation system, and for catalase were measured in isolated hepatocyte nuclei obtained from male rats following a single intragastric dose of ciprofibrate, clofibrate, or (EtHx)2 greater than Pht. All three peroxisome proliferators rapidly increased the rate of FAOxase and PBE gene transcription in liver, with near maximal rates (9-15 times control) reached by 1 hr and persisting until at least 16 hr after administration of the compound. FAOxase and PBE mRNA levels, measured by blot-hybridization analysis and FAOxase and PBE protein content, analyzed by immunoblotting, increased concurrently up to at least 16 hr following a single dose of peroxisome proliferator. The catalase mRNA level increased about 1.4-fold, but the transcription rate of the catalase gene was not significantly affected. The results show that the peroxisome proliferators clofibrate, ciprofibrate, and (EtHx)2 greater than Pht selectively increase the rate of transcription of peroxisomal fatty acid beta-oxidation enzyme genes. Whether the transcriptional effects are mediated by peroxisome proliferator-receptor complexes remains to be elucidated.

32P-postlabeling analysis of peroxisome proliferator-DNA adduct formation in rat liver in vivo and hepatocytes in vitro.

Hepatocarcinogenic peroxisome proliferators, clofibrate, ciprofibrate, Wy-14643 or di(2-ethylhexyl)phthalate, were administered once daily by gavage to groups of three male F344 rats for 3 days and the rats were killed 2 h after the last dose. The DNA isolated from the livers was analyzed for possible carcinogen-DNA adducts, by a most sensitive 32P-postlabeling technique which can detect one adduct in 10(10) nucleotides. No adducts were detected by this assay in the DNA isolated from the livers of rats treated with any of the peroxisome proliferators. Adducts were also not found in the DNA of hepatocytes exposed in vitro to these peroxisome proliferators for 4 h in primary suspension cultures. Failure to detect peroxisome proliferator DNA adducts in hepatocytes under in vivo and in vitro conditions supports the contention that formation of a peroxisome proliferator-DNA adduct is not an essential step in the carcinogenesis by this novel class of carcinogens.