Vitamin D: A Critical Micronutrient for Vascular Health in Type 2 Diabetes Mellitus.

OBJECTIVE: Vitamin D deficiency may be responsible for endothelial dysfunction which in turn affects the onset and progression of vascular disease including coronary artery disease and its risk factor, directly or indirectly through various mechanisms. This study was undertaken to find out association between vitamin D and endothelial dilatation of brachial artery, which may help to suggest possible underlying mechanisms and may be of clinical importance in planning preventive and therapeutic strategies. MATERIAL AND METHODS: 50 cases of type 2 diabetes mellitus aged 40-60 years were taken. 50 cases without type 2 diabetes mellitus matched for confounding factors were taken as controls. Venous blood samples were collected for the investigations including vitamin D levels. Then Participants were subjected to ultrasound examination for measurement of flow mediated dilatation (FMD) and endothelial independent dilatation after ingestion of glyceryl tri-nitrate (GTN). Unpaired student T test and correlation coefficient analysis were used to find out association between different variables. RESULTS: The mean values of FMD were 18.85 ± 5.39% and 10.29 ± 4.91% in controls and cases respectively (p<0.001). The dilatation after GTN was observed to be 26.16 ± 4.25% and 18.74 ± 5.72% in controls and cases respectively (p<0.001). The mean levels of vitamin D among controls and cases were 25.41 ± 12.18 and 14.52 ± 8.28 ng/ml respectively. The correlation between endothelial dependent dilatation (FMD), endothelial independent dilatation (after GTN) and vitamin D was found to be more positive in cases (r=0.870, r=0.798) than controls (r=0.079, r=0.158). CONCLUSIONS: Vitamin D deficiency state is higher among cases of type 2 DM. Endothelial dependent dilatation (FMD) was found to be lower among the patients of type 2 DM. The study gives us an insight to identify the diabetics with vitamin D deficiency which may be at higher risk of vascular complications including coronary artery disease.

Treatment of wastewater and restoration of aquatic systems through an eco-technology based constructed treatment wetlands - a successful experience in Central India.

In the last couple of decades constructed wetlands (CWs) have drawn considerable interest in Central India. CWs offer an effective means of integrating wastewater treatment and resource enhancement, often at competitive cost in comparison to conventional wastewater treatments, with additional benefits of Green Urban Landscaping and wildlife habitat. This paper describes treatment performances and the design of some Sub Surface Flow CWs (SSFCW) and Artificial Floating Islands (AFIs) in Central India. Central Indian CWs show significant pollution reduction load for total suspended solids (TSS) (62-82%), biochemical oxygen demand (BOD) (40-75%), NH(4)-N (67-78%) and total Kjeldahl nitrogen (TKN) (59-78%). Field scale SSFCWs installed so far in Central India are rectangular, earthen, single/multiple celled having similar depths of 0.60-0.90 m, hydraulic retention capacity 18-221 m(3) with effective size 41.8-1,050 m(2). The major components of CWs incorporate puddled bottom/side walls, sealed with impermeable low-density polyethylene, a bed of locally available river gravel planted with Phragmites karka, and an inlet distribution and outlet collection system. A new variant on CWs are AFIs working under hydroponics. The field scale experimental AFIs installed in-situ in a slowly flowing local river were composed of hollow bamboo, a bed of coconut coir, floating arrangements and Phragmites karka as nutrient stripping plant species. The AFIs polish the aquatic system by reducing 46.6% of TSS, 45-55% of NH(4)-N, 33-45% of NO(3)-N, 45-50% of TKN and 40-50% of BOD. The study established that there is a need for further research and sufficient data to assist the development of CWs by instilling confidence in policymakers, planners and in the public.

Outcome of Canal Wall Down Mastoidectomy: A Retrospective Review.

A canal wall down mastoidectomy (CWDM) is an effective technique for eradication of chronic suppurative otitis media (atticoantral disease) or cholesteatomas. A retrospective study was conducted at R.D. Gardi Medical college between 2015 and 2017 to evaluate the outcome of CWDM for patients with atticoantral type of CSOM. All new cases of MRM/RM which fulfilled the selection criteria were reviewed. The main outcome measures were the hearing outcome and status of dry ear postoperatively. 60 patients of CWDM (32 male, 28 female) were included for analysis. All patients in our study were between 7 and 52 years with mean age 28.38 years. The ossicular chain was eroded in all the cases. Improvement in Air-Bone-Gap was seen in 11 (18.33%), no change in 38 (63.33%) and 11 (18.33%) had deteriorated. In post operative outcome study of our 60 patients, 76.66% of the patients had dry healed cavity and non healed cavity was found to be in 23.33% of the patients. On analysis of data and comparison with other studies it was found that there are three important factors which influence hearing results and the outcome of surgery-status of ossicular chain, presence or absence of cholesteatoma, recurrence or residual disease.

Oxidation of polyaromatic hydrocarbons in systems containing water miscible organic solvents by the lignin peroxidase of Gleophyllum striatum MTCC-1117.

Lignin peroxidase has been purified to homogeneity using a process of concentration by ultrafiltration and anion exchange chromatography on diethylaminoethyl (DEAE) cellulose from the liquid culture filtrate of the brown rot fungi Gleophyllum striatum MTCC-1117. The molecular mass of the purified enzyme is 43 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The K(m) values for the enzyme using veratryl alcohol, hydrogen peroxide and n-propanol were 66 microM, 82 microM and 476 microM, respectively. The pH and temperature optima of the enzyme were 2.8 and 25 degrees C, respectively. The enzyme is completely inhibited by 20% of the water miscible organic solvents acetone dioxane, diethylether, acetonitrile and dimethylformamide. The lignin peroxidase oxidizes polycyclic aromatic hydrocarbons pyrene, acenaphthene, anthracene, dibenothiophene and 9-methyl anthracene.

Purification and characterisation of lignin peroxidase from Pycnoporus sanguineus MTCC-137.

Extracellular secretion of lignin peroxidase from Pycnoporus sanguineus MTCC-137 in the liquid culture growth medium amended with lignin containing natural sources has been shown. The maximum secretion of lignin peroxidase has been found in the presence of saw dust. The enzyme has been purified to homogeneity from the culture filtrate of the fungus using ultrafiltration and anion exchange chromatography on DEAE-cellulose. The purified lignin peroxidase gave a single protein band in sodium dodecylsulphate polyacrylamide gel electrophoresis corresponding to the molecular mass 40 kDa. The K(m)(, kcat) and k(cat)/K(m) values of the enzyme using veratryl alcohol and H2O2 as the substrate were 61 microM, 2.13 s(-1), 3.5 x 10(4) M(-1) s(-1) and 71 microM, 2.13 s(-1), 3.0 x 10(4) M(-1) s(-1) respectively at the optimum pH of 2.5. The temperature optimum of the enzyme was 25 degrees C.

Polymorphism of glutathione S-transferase M1 and T1 gene loci in COPD.

Chronic obstructive pulmonary disease (COPD) is a complex combination of signs and symptoms in patients with chronic bronchitis and emphysema, diseases that largely result from cigarette smoking. A little information is available for the underlying molecular mechanisms that are responsible for its occurrence. Polymorphisms in genes of xenobiotics metabolizing enzymes are expected to modulate individual responses to genotoxic carcinogens. Present study was a case-control study of COPD patients and healthy controls. Genetic polymorphisms of GSTM1 and GSTT1 genes in 50 COPD patients and 50 healthy controls were investigated using multiplex polymerase chain reaction-restriction fragment length polymorphism techniques to determine whether polymorphisms of these genes are linked to genetic susceptibility to COPD. All subjects were males and smokers. The frequency of GSTM1 homozygous null genotype was 28.0% in COPD cases when compared with controls (32.0%). The difference was not significant showing that risk of COPD was not associated with the GSTM1 null genotypes. The frequencies of homozygous null genotypes of GSTT1 were significantly higher in COPD cases as compared with controls (40% versus 14.0%) suggesting that the theta-glutathione S-transferases null genotype may be associated with the susceptibility to COPD. No significant differences were observed when comparisons were performed according to severity of disease and smoking for GSTM1 and GSTT1. It was also observed that COPD developed in the early age and with a shorter pack-year history in Indian population.

Parasitic zoonoses in India: an overview.

Parasitic zoonotic diseases are prevalent throughout India at varying rates. First reports of zoonotic parasites and new emerging diseases have been recorded in both the human and animal populations in recent decades. The prevalence of zoonotic parasites is likely to be an underestimate, owing to the lack of proper surveillance and the shortage of information about the existence of asymptomatic animal carriers. Emergence of diseases such as human echinococcosis/hydatidosis, neurocysticercosis, cryptosporidiosis and toxoplasmosis in those with acquired immune deficiency syndrome, together with the re-emergence of cutaneous leishmaniosis, poses a serious threat in India and the prevention and control of these parasitic zoonoses, and others, is a great challenge.

Prevalence of high-risk human papillomavirus type 16/18 infection among women with normal cytology: risk factor analysis and implications for screening and prophylaxis.

OBJECTIVE: To determine the prevalence of high-risk human papillomavirus (HR-HPV) 16/18 infection of uterine cervix among women in the reproductive age group, with cytologically normal cervical (Pap) smears; to analyse the risk factors for HR-HPV acquisition and to address their implications for cervical cancer screening and prophylaxis in a low resource setting. METHODS: Cervical samples from 769 cytologically negative women (age 18-45 years) attending a tertiary care centre in Delhi were subjected to HPV DNA testing and HR-HPV 16/18 and low-risk (LR)-HPV 6/11 sub-typing by polymerase chain reaction. Univariate risk factor analysis was carried out in HR-HPV positive (n = 86) versus HR-HPV negative women (n = 683) by chi-square test. RESULTS: The overall HPV prevalence among cytologically normal women was 16.6%. HR-HPV16 was detected in 10.1%, whereas HPV18 was detected in 1% of women. HR-HPV 16/18 comprised 67% of the total HPV positives. There was no decline in HR-HPV positivity with age, and women aged 40-44 years were at significantly increased risk for HR-HPV prevalence (P = 0.03). Statistically significant associations of HR-HPV infection were found with risk factors such as high parity (P = 0.04), cervicitis/hypertrophic cervix (P = 0.01), unhealthy cervix (P = 0.04), rural residence (P = 0.03), low socioeconomic status (P = 0.01) and illiteracy (P = 0.07). CONCLUSIONS: Although the sample size was small, based on the observation that HR-HPV 16 and 18 contributed significantly to the overall HPV prevalence in our setting, we speculate that testing/prophylaxis for these prevalent high-risk types could perhaps make cervical cancer screening and preventive programmes cost-effective. Larger community-based studies on HPV prevalence and persistence are required to validate these findings before definitive recommendations can be made to the policy makers.

Treatment performance of artificial floating reed beds in an experimental mesocosm to improve the water quality of river Kshipra.

The discharge of untreated wastewater in River Kshipra had brought annual average of BOD, TKN and TS levels up to 39 mg/l, 38 mg/l and 781 mg/l respectively in the study area. Treatment performance by Artificial Floating Reed Beds (AFRB) was evaluated for removal efficiency of TS, NH4-N, NO3-N, TKN and BOD from river water, initially, under a pilot scale by an AFRB of size 200 m2 planted with local reed grass, Phragmites karka, in the part of River Kshipra at the confluence with meeting point of a wastewater stream. The system performance was recorded as 43% reduction in TS, 38% reduction in TKN and 39% BOD reduction. The experimental AFRBs were buoyant structure planted with reed grass, each unit had a rectangular size and covered an effective surface area of 2 m2. The experiment with the mesocosms with treatment of River water resulted that AFRB was reducing pollution load by 55-60% of TS, 45-55% of NH4-N, 33-45% of NO3-N, 45-50% of TKN and 40-50% of BOD. AFRB may be recommended as an in-situ, eco-friendly river water treatment structures for small shallow, slow flowing (or slightly stagnant) water bodies.

Solitary Plasmacytoma of Frontal Bone: A Case Report.

A 42 years old male presenting with complaints of swelling on the left supraorbital region since-5 months. Swelling was gradual in onset and progressive in nature. On examination it was a single swelling 5 × 6 cm, with diffuse margins, skin over the swelling was red and stretched, swelling is firm in consistency, fixed, tender on touch, non pulsatile with diffuse irregular margin. FNAC. Smears are positive for malignancy. Bone marrow examination was normal. Excision of the mass from the left frontal sinus, orbit and supraorbital region under GA and sent for HPR. Definitive treatment of choice is: radiotherary 45-50 Gy/4-5 weeks, chemotherapy and surgery. Prognosis is good with local recurrence (< 10%).

Genetic polymorphism at GSTM1 and GSTT1 gene loci and susceptibility to oral cancer.

GSTs are phase II enzymes which are involved in the detoxification of active metabolites of many potential carcinogens from tobacco smoke and therefore may play an important role in modulating susceptibility to tobacco related cancers. This study evaluates the influence of genetic polymorphisms of GSTM1 and GSTT1 gene loci on susceptibility to oral cancer. The genotyping was based on multiplex PCR assay that identified the GSTM1 and GSTT1 null (-/-) genotypes but didn't distinguish homozygous wild type+/+ and heterozygous +/- individuals. Genomic DNA was isolated from cases with oral cancer (n=40) and normal controls (n=87). The prevalence of the GSTM1 null genotypes was 29/87 (33.3%) and 21/40 (52.5%) in controls and oral cancer cases, respectively but the differences were not significant (OR=2.2; 95%CI=0.96-5.1; p=0.06). The frequency of homozygous GSTT1 null genotype in cancer cases was 17/40 (42.5%) as compared to 13/87 (14.94%) in controls and the differences were highly significant (OR=4.2; 95%CI=1.64-10.9; p=0.0002). Oral cancer cases had higher proportion of both GSTM1 and GSTT1 null genotypes as compared to controls but the differences were not statistically significant (OR=2.9; 95%CI=0.71-11.9; p=0.17). When individuals were categorized into two groups, no differences were observed for GSTM1 null genotype frequencies in control and cancer cases (OR=2.9; 95%CI=0.9-9.6; p=0.08) (OR=1.6; 95%CI=0.44-6.1; p=0.58) in <=50 yrs and >50 yrs of age groups. Significant differences between control and cancer cases were observed for GSTT1 null genotypes both in <=50 yrs and >50 yrs of age groups (OR=4.0; 95%CI=1.1-15.0; p=0.03) (OR=4.5; 95%CI=0.97-22.29; p=0.05), respectively. The effect of smoking on GSTM1 null individuals was not found significant (OR=1.0; 95%CI=0.19-4.86; p=0.75) but it was significant in case of GSTT1 null individuals (OR=6.33; 95%CI=1.0-44.1; p=0.02). Our results thus suggest that GSTT1 gene polymorphisms modulate susceptibility to tobacco-related cancer of the oral cavity.

Histopathological changes associated with E. granulosus echinococcosis in food producing animals in Punjab (India).

Cystic echinococcosis due to Echinococcus granulosus is a serious public health and economic concern in India. The disease is endemic in most of the food producing animals such as cattle, buffalo, sheep, goat and pigs in the country. In this study, tissues comprising of pieces of liver and lungs were collected in 10 % formal buffered saline. The formalin fixed tissues (liver and lungs) from 10 cattle, buffalo, sheep, goat and pigs each were selected and further processed by acetone benzene method for histopathological examinations. The cysts were surrounded by outer fibrous layer over the inner germinal layer and filled with clear hydatid fluid. Echinococcal protoscolices were also noticed in some of the sections. Histologically, slight hemorrhage, leucocyte infiltration and mild hepatocellular degeneration in the liver were noticed. The adjacent hepatic paraenchyma showed atrophy, variable degeneration and infiltration. The parenchyma adjacent to cysts was markedly congested and showed multiple small haemorrhagic areas. In lungs, there was proliferation of fibrous connective tissue and infiltration of mononuclear cells.

Techniques in Cardiology : PDA Closure in Children.

BACKGROUND: Transcatheter closure of patent ductus arteriosus (PDA) with various devices has been evaluated worldwide and in selected cases can be performed successfully, thus avoiding the morbidity associated with surgical closure. METHODS AND RESULTS: A 2 year old female child presented with PDA and aortic angiography showed large PDA (tubular). This defect can be closed nonsurgically by coils and devices. The coil used are normally 0.038" having delivery system called Flipper, but this child had a large PDA and so was closed by thicker coil 0.052" using a coil-Bioptome-sheath system, which is the first reported case from the Armed Forces. CONCLUSIONS: We report the initial experience at our center of closure of PDA with a new coil-Bioptome-sheath system.

Prevalence and morphological characterisation of Cysticercus tenuicollis (Taenia hydatigena cysts) in sheep and goat from north India.

Taenia hydatigena is an adult parasite of dogs with the metacestode (Cysticercus tenuicollis) stage residing in ruminants and pigs. Documentation and surveillance data concerning to the prevalence and risk factors associated with the disease in India is largely lacking. In this experiment, 3,199 carcasses, including 760 sheep and 2,439 goat were examined for the presence of C. tenuicollis (T. hydatigena cysts) on post-mortem inspection at different slaughter houses/shops in northern India. Morphological analysis was also conducted on five samples from each species. Out of 3199 carcasses examined, 135 were found containing cysts of T. hydatigena indicating a prevalence of 4.22 %. Most of the cysts were present in abdominal cavity, except few which were embedded in the liver. The high prevalence of 4.83 was recorded in goats as compared to 2.23 % in sheep. Principal component analysis was applied for statistical analysis. The results of morphological analysis indicated its usefulness as a valid criterion for differentiation of T. hydatigena cysts and that there might be possibility of two different strains infecting sheep and goat.

Sister chromatid exchanges in the lymphocytes of patients with oral leukoplakia.

The incidence of sister chromatid exchange (SCE) was investigated in lymphocyte chromosomes of 59 patients with oral leukoplakia and 65 age- and sex-matched nonsmoking controls. The frequency of SCE was found to be 8.61 +/- 1.89 in patients with oral leukoplakia, which was significantly higher than the mean SCE value of 5.58 +/- 1.26 observed in normal controls. The frequency of SCE in patients with oral leukoplakia addicted to the single habit of betel with tobacco chewing, bidi/cigarette smoking, and combined habits of chewing and smoking of tobacco were found to be 7.95 +/- 1.63, 8.17 +/- 1.66, and 9.23 +/- 2.14, respectively. These values were also significantly higher as compared to the SCE values observed in normal controls.

Nucleolar organizer regions in patients with precancerous and cancerous lesions of the uterine cervix.

Nucleolar organizer regions (Ag-NORs) were studied in patients with cervical precancerous and cancerous lesions along with controls. The data revealed a statistically significant decrease of Ag-NORs in cancer patients, as well as in women with severe precancerous lesions as compared with controls. A similar decrease in the amount of Ag-staining was also observed in both cancer and severe dysplasia cases. The study suggests a possible relationship of Ag-NOR activity to malignancy.

A simple and rapid method of high quantity DNA isolation from cervical scrapes for detection of human papillomavirus infection.

Infection with human papillomavirus (HPV) is an important etiological factor in the development of cervical cancer, and detection of the viral genome is of prognostic importance, particularly for preneoplastic lesions. We developed a simple, easy and efficient non-organic method of DNA extraction from cervical scrapes for reliable detection of HPV DNA sequences. The method involves incubation of cell nuclei in higher concentration of proteinase K at 65 degrees C for 2.5 h. Following prolonged incubation at higher temperature, the enzyme is autoinactivated and the DNA isolated can be used directly for analysis without further purification. The recovery of DNA is more than 95% and it can be easily cleaved by restriction enzymes and is suitable for amplification by the polymerase chain reaction (PCR). The whole procedure is carried out in a single Eppendorf tube and a large number of specimens can be processed at a time without any error of handling. DNA extracted from a single smear sample is sufficient to conduct as many as four different molecular biology tests. This provides an opportunity for verification of sensitivity, specificity and reliability of each test for diagnosis of HPV infection without resorting to biopsy.

Hepatitis C virus infection in sporadic fulminant viral hepatitis in North India: cause or co-factor?

INTRODUCTION: The role of hepatitis C virus (HCV) infection in fulminant hepatitis (FH) is poorly understood and the available data are conflicting. We have examined the aetiological role of HCV in 50 consecutive patients with sporadic FH by employing serology and reverse transcription-polymerase chain reaction (RT-PCR). MATERIALS AND METHODS: A total of 50 consecutive patients with sporadic FH were included. After an initial clinical and biochemical assessment, tests were performed for detection of HBsAg, IgM anti-HBc, IgM anti-HAV, IgM anti-HEV and anti-HCV. RT-PCR was carried out for detection of HCV RNA in sera of all the patients and in post mortem liver biopsy tissue of 20 subjects, using primers selected from the conserved 5' non-coding region of the HCV genome. RESULTS: Hepatitis E virus (HEV) was found to be the most common viral infection (21/50; 42%) followed by HBV (14/50; 28%), HCV (7/50; 14%) and HAV (2/50; 4%). No viral markers could be detected in nine patients (18%) and multiple infections were seen in seven (14%). Of the seven subjects who tested positive for HCV-related markers, two had both anti-HCV and HCV RNA, three had HCV RNA alone and the remaining two had anti-HCV alone. Interestingly, all the HCV-infected subjects were co-infected with other hepatotropic viruses and the most common co-infecting agent was found to be HBV (5/7). Liver tissue was available in 20 cases and HCV RNA was detected in three of them. All of these patients were also positive for the viral genome in their serum samples. Comparison of the biological attributes of HCV-positive and HCV-negative cases revealed that haemorrhagic symptomatology (haematemesis, melaena and purpurae) was significantly more common, prothrombin time more deranged and mortality was much higher in the former group. The overall mortality was 68% and the most common cause of death was cerebral oedema (70.6%). No significant correlation was observed between mortality and the duration of the icterus-encephalopathy interval. The study included a total of 21 pregnant females; HEV infection was found to be significantly greater in this group and was associated with a higher mortality rate. CONCLUSIONS: The results clearly suggest that HCV is not an important aetiological factor for FH in North India. However, it may act as a co-factor in the development of FH leading to a higher mortality. HEV appears to contribute substantially to the causation of sporadic FH in India and advanced stage pregnancy is a potential risk factor for HEV-induced FH and high rate of mortality. Our study also suggests that the length of the icterus-encephalopathy period may not have significant prognostic implications in Indian patients with FH.

Development of an avidin-biotin dot enzyme-linked immunosorbent assay and its comparison with other serological tests for diagnosis of glanders in equines.

A dot enzyme-linked immunosorbent assay (dot ELISA) was developed for diagnosis of glanders in equines. The test was based on the detection of IgG antibodies to Pseudomonas mallei antigens bound to nitrocellulose coated on plastic strips (dipsticks), the reaction being amplified by an avidin-biotin system with biotinylated anti-horse IgG and horseradish peroxidase-avidin D. Sera from 810 normal, six naturally infected and 48 sensitized equines were tested by this assay, and results were compared with complement fixation, indirect haemagglutination and counter-immunoelectrophoresis tests. Dot ELISA had the highest sensitivity, and was superior to other tests in that it was rapid and easy to perform, the results were easy to interpret, the assay was not influenced by anti-complement activity, and it was able to detect antibodies at an early stage. Testing of serum at 1:200 dilution is proposed for epidemiological screening.

Potency of partially purified malleo-proteins for mallein test in the diagnosis of glanders in equines.

Malleo-proteins from synthetic broth mallein of six strains of Pseudomonas mallei were separated by trichloroacetic acid precipitation, ammonium sulfate precipitation and Ultrogel AcA 34 gel filtration chromatography. When tested comparatively with Dutch PPD mallein as standard on P. mallei-sensitized and normal horses all the strains were found to be malleinogenic, trichloroacetic acid precipitated proteins were comparable to Dutch PPD mallein in potency and innocuity whereas ammonium sulfate-precipitated proteins elicited non-specific reactions. Ultrogel AcA 34 chromatographed high molecular weight proteins (MW range > 350,000) were having equal or higher potency and without adverse reactions and low molecular weight proteins (MW range 120,000-170,000) nearly inactive. Ultrogel AcA 34 column fractionation studies revealed that mallein activity was associated with high molecular weight proteins and represented a purer sensitin than PPD mallein for the purpose of mallein test.