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1.
J Family Med Prim Care ; 12(9): 2090-2096, 2023 Sep.
Article in English | MEDLINE | ID: mdl-38024948

ABSTRACT

Introduction: Iodine deficiency disorders (IDD) have remained an unresolved public health problem in India. In this survey, we have estimated the prevalence of IDD among 6-12 years of school children in rural areas of north Karnataka, India and estimated the prevalence of low iodine content (<15 ppm) in salt at the household level and urine iodine excretion in this population. Material and Methods: In this cross-sectional survey, we recruited 16,827 children between 6 and 12 years of age through multistage sampling from six districts. Goitre was examined clinically for all children. Household-level salt iodine estimation and urinary iodine estimation were carried out among a subset of the participants. Results: Overall prevalence of goitre was 17.1% (95% CI: 16.5, 17.7). Out of this, 76.7% (n = 2116) had Grade-1 goitre, and 23.7% (n = 656) had Grade-2 goitre. The prevalence of goitre was higher among females (17.9%, vs. male 16.4%, P < 0.05). The prevalence of low iodine content (<15 ppm) in salt was 48.5% (95%CI: 46.7, 50.3). The overall median iodine excretion in urine was 85 µg/L (IQR: 60-150 µg/L). In total, 37.2% (n = 601) had mild iodine deficiency, 5.2% (n = 84) had moderate deficiency, and 10.1% (n = 163) had severe deficiency. All parameters showed high inter- and intradistrict variations. Conclusion: North Karnataka has a high goitre prevalence. Low use of iodized salt can be a major reason for the high prevalence of the condition. Ensuring the availability of iodized salt in this region and periodic surveillance to measure the impact of the programme should be the priority in this region.

2.
Front Nutr ; 10: 1209949, 2023.
Article in English | MEDLINE | ID: mdl-37502723

ABSTRACT

Background: For school-age children, a healthy transition from childhood to adolescence and adulthood depends on proper nutrition. Globally, most nutritional surveys focus on preschool and adolescents, neglecting school-age children. Recent studies have shown the prevalence of thinness among adolescents to be 26.5% in Karnataka. Similarly, among children aged < 5 years in the Raichur district, the prevalence of stunting, wasting, and being underweight was 39.8, 23.2, and 40.7%, respectively. The present study aimed to bridge the data gap between < 5 years of children and adolescents through a nutritional survey of school-going children in Raichur, one of the aspirational districts of India. Materials and methods: A cross-sectional survey was conducted from January to March 2020 among rural school-age children (n = 2700) in 30 villages of the Raichur district of Karnataka, India. The school children were selected through a multi-stage cluster sampling technique. The WHO Anthro-plus software was used for calculating the age and sex-specific Z-scores for weight-for-age (WAZ), height-for-age (HAZ), and BMI-for-age (BAZ). Results: Of the 2,700 school-age children surveyed, the mean weight and height were 22.2 kg (+5.8) and 124.9 cm (+11.6), respectively. The prevalence of children having weight-for-age Z-scores < -2 SD (Underweight) was 45.3% (95% CI: 42.7%-47.8%). The magnitude of stunting and severe stunting was 19.5% (95% CI: 18.0%-21.0%) and 7.6% (95% CI: 6.6%-8.6%), respectively. The proportion of children with BMI for age Z-scores < -2SD (thinness) was 43% (95% CI: 41.1%-44.9%), with sub-district Sindhanur having a dual burden of malnutrition. Conclusion: Despite many flagship programs, the prevalence of undernutrition in school-age children remains a considerable public health problem in the aspirational district of Raichur, India. Furthermore, exploratory studies are recommended to identify the factors associated with undernutrition among school-age children and strategize evidence-based intervention.

3.
Front Public Health ; 11: 1110777, 2023.
Article in English | MEDLINE | ID: mdl-37006577

ABSTRACT

Objectives: The present study was planned to estimate the prevalence of dental fluorosis in 6-12 years of children and its association with various drinking water sources, water, and urine fluoride levels among the subset of children under the umbrella of a larger study to address iodine deficiency disorders and iron deficiency anemia in 17 villages of Manvi and Devadurga talukas of Raichur district of Karnataka. Methods: Analysis of subset of data and urine samples of children under the umbrella of a larger cross-sectional community-based study was conducted in 17 villages of Manvi and Devadurga taluks of Raichur district. House to house survey was carried out to collect data using a semi-structured questionnaire in ODK software. Demographic details, source of drinking water, clinical assessment of dental fluorosis, and height and weight measurements were performed by trained staff. Urine and water samples were collected for fluoride level estimation. The overall prevalence of dental fluorosis and its severity-wise prevalence were estimated. Association between dental fluorosis and age, gender, type of diet, source of drinking water, height for age, BMI for age, water fluoride level, and urine fluoride level were carried out using logistic regression analysis. Results: The prevalence of dental fluorosis was 46.0%. Mild, moderate, and severe dental fluorosis was found in 37.9, 7.8, and 0.3% of children. With the increasing age of participants, the odds of dental fluorosis were found to increase by 2-4 folds. The odds of having dental fluorosis were significantly increased with increasing water fluoride levels of 3 to 5 ppm [AOR = 3.147 (1.585-6.248); P = 0.001] in comparison with water fluoride levels of < 1 ppm. The similar trend was found with urine fluoride level > 4 ppm [AOR = 3.607 (1.861-6.990); P < 0.001]. As compared to river water, other sources of drinking water were significantly associated with higher odds of dental fluorosis. Conclusions: Prevalence of dental fluorosis was high in 6 to 12 years due to overexposure of fluoride from drinking water. High water and urine fluoride levels in children indicate the chronic exposure to fluoride and suggest that the population is at high risk of developing chronic fluorosis.


Subject(s)
Drinking Water , Fluorosis, Dental , Humans , Child , Fluorides/analysis , Fluorosis, Dental/epidemiology , Cross-Sectional Studies , India/epidemiology
4.
J Neonatal Perinatal Med ; 14(3): 403-409, 2021.
Article in English | MEDLINE | ID: mdl-33459668

ABSTRACT

OBJECTIVE: Identify perinatal risk factors associated with SIPSTUDY DESIGN:This was a retrospective case-control study of SIP in infants born at ≤28 weeks of gestation and admitted between 1995 and 2016 at a tertiary care NICU. Infants with NEC or other GI abnormalities were excluded. Cases of SIP were matched with gestational age-matched controls with the closest birth date. Maternal, infant and birth related factors were evaluated using univariate analyses and significant factors were evaluated using multiple logistic regression. RESULT: 25 cases of SIP were matched with 25 controls. No maternal factors reached statistical significance. Being one of twins increased the odds of SIP 29-fold. Birth-order or weight-discrepancy in twin had no association of SIP within twin pairs. CONCLUSION: Twins are at significantly higher risk for SIP. The association of SIP and twin gestation was independent of previously reported risk factors of perinatal indomethacin or magnesium sulfate and merits further study.


Subject(s)
Infant, Premature, Diseases , Intestinal Perforation , Case-Control Studies , Female , Humans , Infant , Infant, Newborn , Infant, Premature , Intestinal Perforation/etiology , Pregnancy , Retrospective Studies
5.
PLoS Comput Biol ; 16(4): e1007750, 2020 04.
Article in English | MEDLINE | ID: mdl-32251432

ABSTRACT

In multicellular organisms, the timing and placement of gene expression in a developing tissue assigns the fate of each cell in the embryo in order for a uniform field of cells to differentiate into a reproducible pattern of organs and tissues. This positional information is often achieved through the action of spatial gradients of morphogens. Spatial patterns of gene expression are paradoxically robust to variations in morphogen dosage, given that, by definition, gene expression must be sensitive to morphogen concentration. In this work we investigate the robustness of the Dorsal/NF-κB signaling module with respect to perturbations to the dosage of maternally-expressed dorsal mRNA. The Dorsal morphogen gradient patterns the dorsal-ventral axis of the early Drosophila embryo, and we found that an empirical description of the Dorsal gradient is highly sensitive to maternal dorsal dosage. In contrast, we found experimentally that gene expression patterns are highly robust. Although the components of this signaling module have been characterized in detail, how their function is integrated to produce robust gene expression patterns to variations in the dorsal maternal dosage is still unclear. Therefore, we analyzed a mechanistic model of the Dorsal signaling module and found that Cactus, a cytoplasmic inhibitor for Dorsal, must be present in the nucleus for the system to be robust. Furthermore, active Toll, the receptor that dissociates Cactus from Dorsal, must be saturated. Finally, the vast majority of robust descriptions of the system require facilitated diffusion of Dorsal by Cactus. Each of these three recently-discovered mechanisms of the Dorsal module are critical for robustness. These mechanisms synergistically contribute to changing the amplitude and shape of the active Dorsal gradient, which is required for robust gene expression. Our work highlights the need for quantitative understanding of biophysical mechanisms of morphogen gradients in order to understand emergent phenotypes, such as robustness.


Subject(s)
Body Patterning/genetics , Drosophila Proteins/genetics , Nuclear Proteins/genetics , Phosphoproteins/genetics , Transcription Factors/genetics , Animals , Cell Nucleus/metabolism , Cytoplasm/metabolism , DNA-Binding Proteins/metabolism , Drosophila/embryology , Drosophila/genetics , Drosophila Proteins/metabolism , Embryo, Nonmammalian/metabolism , Gene Expression Regulation, Developmental/genetics , Morphogenesis/genetics , NF-kappa B/metabolism , Nuclear Proteins/metabolism , Phosphoproteins/metabolism , Signal Transduction , Transcription Factors/metabolism
6.
Curr Top Dev Biol ; 137: 143-191, 2020.
Article in English | MEDLINE | ID: mdl-32143742

ABSTRACT

The morphogen gradient of the transcription factor Dorsal in the early Drosophila embryo has become one of the most widely studied tissue patterning systems. Dorsal is a Drosophila homolog of mammalian NF-κB and patterns the dorsal-ventral axis of the blastoderm embryo into several tissue types by spatially regulating upwards of 100 zygotic genes. Recent studies using fluorescence microscopy and live imaging have quantified the Dorsal gradient and its target genes, which has paved the way for mechanistic modeling of the gradient. In this review, we describe the mechanisms behind the initiation of the Dorsal gradient and its regulation of target genes. The main focus of the review is a discussion of quantitative and computational studies of the Dl gradient system, including regulation of the Dl gradient. We conclude with a discussion of potential future directions.


Subject(s)
Body Patterning , Computational Biology/methods , Drosophila Proteins/metabolism , Drosophila melanogaster/physiology , Embryo, Nonmammalian/physiology , Gene Expression Regulation, Developmental , NF-kappa B/metabolism , Nuclear Proteins/metabolism , Phosphoproteins/metabolism , Transcription Factors/metabolism , Animals , Drosophila Proteins/genetics , Drosophila melanogaster/embryology , Embryo, Nonmammalian/cytology , NF-kappa B/genetics , Nuclear Proteins/genetics , Phosphoproteins/genetics , Signal Transduction , Transcription Factors/genetics
7.
Dev Dyn ; 249(3): 369-382, 2020 03.
Article in English | MEDLINE | ID: mdl-31925874

ABSTRACT

BACKGROUND: A feedforward loop (FFL) is commonly observed in several biological networks. The FFL network motif has been mostly studied with respect to variation of the input signal in time, with only a few studies of FFL activity in a spatially distributed system such as morphogen-mediated tissue patterning. However, most morphogen gradients also evolve in time. RESULTS: We studied the spatiotemporal behavior of a coherent FFL in two contexts: (a) a generic, oscillating morphogen gradient and (b) the dorsal-ventral patterning of the early Drosophila embryo by a gradient of the NF-κB homolog dorsal with its early target Twist. In both models, we found features in the dynamics of the intermediate node-phase difference and noise filtering-that were largely independent of the parameterization of the models, and thus were functions of the structure of the FFL itself. In the dorsal gradient model, we also found that proper target gene expression was not possible without including the effect of maternal pioneer factor Zelda. CONCLUSIONS: An FFL buffers fluctuation to changes in the morphogen signal ensuring stable gene expression boundaries.


Subject(s)
Body Patterning/physiology , Drosophila Proteins/metabolism , Embryo, Nonmammalian/metabolism , Animals , Body Patterning/genetics , Drosophila Proteins/genetics , Drosophila melanogaster , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Developmental/physiology , NF-kappa B/metabolism , Signal Transduction/genetics , Signal Transduction/physiology
8.
J Endocrinol Invest ; 43(7): 925-934, 2020 Jul.
Article in English | MEDLINE | ID: mdl-31927747

ABSTRACT

BACKGROUND: Thyroid nodules (TN) are detected in a small number of asymptomatic children and adolescents but are more frequently malignant (22-26%) than in adults leading some clinicians to perform thyroid ultrasound (US) for all children with goiter or autoimmune thyroiditis (AIT). Our study was designed to determine if suspicious clinical features predict the presence of TN in children with goiter or AIT so that US could be performed on those at highest risk. METHODS: This was a retrospective review of 223 children and adolescents with goiter or AIT evaluated at a single institution. US was not performed on all patients. It is our practice to define glands that are large, firm, or nodular to palpation as "suspicious". Suspicious glands were interrogated by US and if TN was confirmed, this was further evaluated by fine-needle aspiration followed by surgery if indicated. RESULTS: The median age was 12.9 years with 74.4% female. TN were confirmed by US in 16.6% of all patients but only 4.8% of those with AIT. By univariate analysis, TN were more common in those with family history of TN or differentiated thyroid carcinoma (DTC), thyroid asymmetry, and lower thyrotropin (TSH) levels. Differentiated thyroid carcinoma (DTC) was identified in 10.8% of TN and 1.8% of all patients. Firmness was significantly more common in patients with DTC (p = 0.0013). CONCLUSION: TN were less common in those with AIT than reported in previous studies, suggesting that clinical features might fail to identify the majority of TN in patients with AIT. However, patients with asymmetric thyroid and a family history of TN or DTC have greatest risk for TN.


Subject(s)
Thyroid Nodule/diagnosis , Thyroid Nodule/epidemiology , Adolescent , Age of Onset , Child , Child, Preschool , Cohort Studies , Female , Goiter/diagnosis , Goiter/epidemiology , Goiter/pathology , Humans , Male , Pediatrics/methods , Prognosis , Retrospective Studies , Risk Assessment , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/epidemiology , Thyroid Neoplasms/pathology , Thyroid Nodule/pathology , Thyroiditis, Autoimmune/diagnosis , Thyroiditis, Autoimmune/epidemiology , Thyroiditis, Autoimmune/pathology , Ultrasonography , Virginia/epidemiology
9.
Adv Biomed Res ; 6: 97, 2017.
Article in English | MEDLINE | ID: mdl-28828348

ABSTRACT

BACKGROUND: Staphylococcus aureus has the ability to form biofilms on any niches, a key pathogenic factor of this organism and this phenomenon is directly related to the concentration of NADPH. The formation of NADP is catalyzed by NAD kinase (NADK) and this gene of S. aureus ATCC 12600 was cloned, sequenced, expressed and characterized. MATERIALS AND METHODS: The NADK gene was polymerase chain reaction amplified from the chromosomal DNA of S. aureus ATCC 12600 and cloned in pQE 30 vector, sequenced and expressed in Escherichia coli DH5α. The pure protein was obtained by passing through nickel metal chelate agarose column. The enzyme kinetics of the enzyme and biofilm assay of the S. aureus was carried out in both aerobic and anaerobic conditions. The kinetics was further confirmed by the ability of the substrates to dock to the NADK structure. RESULTS: The recombinant NADK exhibited single band with a molecular weight of 31kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the gene sequence (GenBank: JN645814) revealed presence of only one kind of NADK in all S. aureus strains. The enzyme exhibited very high affinity for NAD compared to adenosine triphosphate concurring with the docking results. A root-mean-square deviation value 14.039Å observed when NADK structure was superimposed with its human counterpart suggesting very low homology. In anaerobic conditions, higher biofilm units were found with decreased NADK activity. CONCLUSION: The results of this study suggest increased NADPH concentration in S. aureus plays a vital role in the biofilm formation and survival of this pathogen in any environmental conditions.

10.
Microb Ecol Health Dis ; 28(1): 1298340, 2017.
Article in English | MEDLINE | ID: mdl-28572751

ABSTRACT

​Background: Consumption of Lactobacillus paracasei Lpc-37 or Bifidobacterium lactis HN019 by 2-5-year-old children was found to reduce risk for diarrhoea and fever during the rainy season. Objective: Can changes in faecal short chain fatty acids (SCFAs) or branched chain fatty acids (BCFAs) explain the observed positive influence of probiotics and their role on nutritional status and diarrhoea risk? Design: Faecal samples were analysed for SCFAs and BCFAs and correlated to Bifidobacterium and Lactobacillus levels; both at the start and after nine months' consumption of either of the two probiotic strains, or placebo. Results: No differences in SCFAs, BCFAs, Lactobacillus or Bifidobacterium levels were found between boys and girls. Severely underweight children were observed to have the highest Lactobacillus levels. Probiotic intervention was found to be associated with higher levels of selected SCFAs and BCFAs in subjects who had experienced diarrhoea. Treatment with either of the probiotics led to changes in SCFAs and BCFAs. SCFAs, acetate, propionate and butyrate, were found to correlate with each other. Likewise, BCFAs isobutyrate, 2-methylbutyrate and isovalerate correlated with each other. After the intervention, L. paracasei Lpc-37 correlated positively with total Bifidobacterium counts and isovalerate levels. B. lactis HN019 counts were found to correlate positively with total bacterial counts and negatively with propionate levels. Conclusions: ​Nutritional status was associated with higher levels of faecal lactobacilli; the meaning of this requires further investigation. The intervention with the two probiotics was observed to influence the levels of faecal SCFAs and BCFAs and there is a differential response in those who developed diarrhoea and those who did not. It is, however, not clear to what extent this is a mechanism that explains the earlier observed effect the strains had on diarrhoea risk.

11.
Indian J Med Res ; 141(1): 46-54, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25857494

ABSTRACT

BACKGROUND & OBJECTIVES: Information on nutritional status of HIV infected children from India is lacking and is required before taking up nutritional supplementation trials. Thus, the aim of the present study was to assess the growth and morbidity status of HIV infected children over a period of one year in a city in southern India. METHODS: This was an observational study carried out between July 2009 and February 2011, at two orphanages in Hyderabad, India. Seventy seven HIV-positive children aged between 1 and half and 15 years, both on and not on antiretroviral therapy (ART) were included. Nutritional status was assessed longitudinally for one year by weight gain, linear growth and body composition. Serum samples were analyzed for haemoglobin, micronutrients, CD4 and CD8 counts. Dietary intakes were assessed by institutional diet survey and morbidity data were recorded every day for 12 months. RESULTS: Mean energy intakes were less than recommended dietary allowance (RDA) in all age groups. Iron and folate intakes were less than 50 per cent of RDA; 46 (59.7%) children were stunted, 36 (46.8%) were underweight and 15 (19.5%) had low BMI for age. Anaemia was observed in 35 (45.5%) children. Micronutrient deficiencies such as vitamin D (40/77; 51.9%), vitamin A (11/77; 14.3%), folate (37/77; 48.1%), iron (38/77; 49.3%) were widely prevalent. HIV viral load was higher in children not on ART and those with morbidity. Respiratory (36.6%) and dermatological illnesses (18.8%) were the commonest presentations. INTERPRETATION & CONCLUSIONS: Acute, chronic malnutrition and micronutrient deficiencies were common in HIV infected children, especially in those not on ART and having morbidity. With severe malnutrition being an alarming consequence of HIV, prophylactic nutritive care should be considered for integration into HIV care strategies besides initiation of ART to improve the nutritional status and quality of life of these children.


Subject(s)
HIV Infections/physiopathology , Health Status , Nutritional Status , Adolescent , Child , Child, Preschool , Female , HIV Infections/blood , Humans , India , Infant , Male
12.
J Enzyme Inhib Med Chem ; 30(4): 655-61, 2015.
Article in English | MEDLINE | ID: mdl-25744510

ABSTRACT

Isocitrate dehydrogenase (IDH) gene from Staphylococcus aureus ATCC12600 was cloned, sequenced and characterized (HM067707). PknB site was observed in the active site of IDH; thus, it was predicted as IDH may be regulated by phosphorylation. Therefore, in this study, PknB, alkaline phosphatase III (SAOV 2675) and IDH genes (JN695616, JN645811 and HM067707) of S. aureus ATCC12600 were over expressed from clones PV 1, UVPALP-3 and UVIDH 1. On passing the cytosloic fractions through nickel metal chelate column, pure enzymes were obtained. Phosphorylation of pure IDH by PknB resulted in the complete loss of activity and was restored upon dephosphorylation with SAOV 2675 which indicated that phosphorylation and dephosphorylation regulate IDH activity in S. aureus. Further, when S. aureus ATCC12600 was grown in BHI broth, decreased IDH activity and increased biofilm units were observed; therefore, this regulation of IDH alters redox status in this pathogen favouring biofilm formation.


Subject(s)
Biofilms , Isocitrate Dehydrogenase/metabolism , Staphylococcus aureus/enzymology , Amino Acid Sequence , Isocitrate Dehydrogenase/chemistry , Isocitrate Dehydrogenase/genetics , Molecular Sequence Data , Phosphorylation , Sequence Homology, Amino Acid
13.
3 Biotech ; 5(4): 505-512, 2015 Aug.
Article in English | MEDLINE | ID: mdl-28324552

ABSTRACT

Staphylococcus aureus, a natural inhabitant of nasopharyngeal tract, survives mainly as biofilms. Previously we have observed that S. aureus ATCC 12600 grown under anaerobic conditions exhibited high rate of biofilm formation and L-lactate dehydrogenase activity. Thus, the concentration of pyruvate plays a critical role in S. aureus, which is primarily catalyzed by pyruvate kinase (PK). Analyses of the PK gene sequence (JN645815) revealed presence of PknB site in PK gene indicating that phosphorylation may be influencing the functioning of PK. To establish this hypothesis the pure enzymes of S. aureus ATCC 12600 were obtained by expressing these genes in PK 1 and PV 1 (JN695616) clones and passing the cytosolic fractions through nickel metal chelate column. The molecular weights of pure recombinant PK and PknB are 63 and 73 kDa, respectively. The enzyme kinetics of pure PK showed K M of 0.69 ± 0.02 µM, while the K M of PknB for stpks (stpks = NLCNIPCSALLSSDITASVNCAK) substrate was 0.720 ± 0.08 mM and 0.380 ± 0.07 mM for autophosphorylation. The phosphorylated PK exhibited 40 % reduced activity (PK = 0.2 ± 0.015 µM NADH/min/ml to P-PK = 0.12 ± 0.01 µM NADH/min/ml). Elevated synthesis of pyruvate kinase was observed in S. aureus ATCC 12600 grown in anaerobic conditions suggesting that the formed pyruvate is more utilized in the synthesis phase, supporting increased rate of biofilm formation.

14.
Biotech Histochem ; 90(1): 8-13, 2015 Jan.
Article in English | MEDLINE | ID: mdl-24867495

ABSTRACT

The incidence of potentially malignant oral pathology such as leukoplakia, oral submucous fibrosis and squamous cell carcinoma has increased in India. We investigated whether cytoplasmic diameter, nuclear diameter and nucleus:cytoplasm ratio in exfoliative cytology are reliable indicators of potentially malignant lesions. We also investigated methyl green-pyronin Y and Feulgen staining as simple time saving and cost effective staining techniques for diagnostic exfoliative cytology. Cell and nuclear diameters of squamous cells of normal buccal mucosa, oral leukoplakia and oral submucous fibrosis were measured using an ocular micrometer disc. The nucleus:cytoplasm ratios in pathological cells were compared to age, sex and site matched controls. We found a significant reduction in the mean cytoplasmic and nuclear diameter in the experimental groups compared to normal controls. Methyl green-pyronin Y stained smears were clearer than Feulgen stained cells. We suggest that a decreased mean cytoplasmic diameter of exfoliated buccal mucosal cells could serve as an early indicator of dysplastic change in lesions that otherwise appear benign. Methyl green-pyronin Y may be useful for identifying premalignant and malignant transformations before a lesion is visible. The simplicity of the technique makes its routine use feasible.


Subject(s)
Leukoplakia, Oral/pathology , Mouth Neoplasms/pathology , Oral Submucous Fibrosis/pathology , Rosaniline Dyes , Cell Nucleus/pathology , Cytodiagnosis/methods , Cytoplasm/pathology , Female , Humans , Leukoplakia, Oral/diagnosis , Male , Methyl Green , Mouth Mucosa/pathology , Mouth Neoplasms/diagnosis , Precancerous Conditions/diagnosis , Pyronine , Staining and Labeling/methods
15.
Indian J Pharm Sci ; 76(5): 430-6, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25425757

ABSTRACT

Glucokinase is classified in bacteria based upon having ATP binding site and 'repressor/open reading frames of unknown function/sugar kinases' motif, the sequence of glucokinase gene (JN645812) of Staphylococcus aureus ATCC12600 showed presence of ATP binding site and 'repressor/open reading frames of unknown function/sugar kinases' motif. We have earlier observed glucokinase of S. aureus has higher affinity towards the substrate compared to other bacterial glucokinase and under anaerobic condition with increased glucose concentration S. aureus exhibited higher rate of biofilm formation. To establish this, 3D structure of glucokinase was built using homology modeling method, the PROCHECK and ProSA-Web analysis indicated this built glucokinase structure was close to the crystal structure. This structure was superimposed with different bacterial glucokinase structures and from the root-mean-square deviation values, it is concluded that S. aureus glucokinase exhibited very close homology with Enterococcus faecalis and Clostridium difficle while with other bacteria it showed high degree of variations both in domain and nondomain regions. Glucose docking results indicated -12.3697 kcal/mol for S. aureus glucokinase compared with other bacterial glucokinase suggesting higher affinity of glucose which correlates with enzyme kinetics and higher rate of biofilm formation.

16.
Appl Biochem Biotechnol ; 169(3): 862-9, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23288593

ABSTRACT

The Krebs cycle dictates oxidative and reductive conditions in Staphylococcus aureus and is mainly regulated by isocitrate dehydrogenase (IDH) which plays pivotal role in the growth and pathogenesis of the bacteria. In the present study, IDH gene from S. aureus ATCC12600 was cloned in the Sma I site of pQE 30 vector; the resultant clone was named as UVIDH1. The insert in the clone was sequenced (accession number HM067707), and the sequence showed complete homology with IDH sequence of other S. aureus strains reported in the database indicating presence of single enzyme in S. aureus, and considerable sequence homology with other bacteria was observed; however, only 24% homology was found with NADP-dependent human IDH. Phylogenetically, the S. aureus IDH showed close identity with Bacillus subtilis and high degree of variability with other bacteria and human IDH. The expression of IDH in the clone UVIDH1 was induced with 1 mM IPTG, and the recombinant IDH was purified by passing through nickel metal chelate column; the purified recombinant IDH showed a single band in SDS-PAGE with a molecular weight of 40 kDa; K(m) and V(max) for isocitrate are 8.2 ± 0.28 and 525 ± 25 µM NADPH/mg/min, respectively, and for cofactor NADP 67.5 ± 2.82 µM and V(max) 50.5 ± 2.12 µM NADPH/mg/min.


Subject(s)
Bacterial Proteins/metabolism , Isocitrate Dehydrogenase/metabolism , Staphylococcus aureus/enzymology , Electrophoresis, Polyacrylamide Gel , Molecular Weight
17.
Indian J Med Res ; 136(6): 963-70, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23391791

ABSTRACT

BACKGROUND & OBJECTIVES: Estimation of parathyroid hormone (PTH) levels is important in the management of metabolic bone disorders. Here we describe a simple, sensitive and specific second generation immunoradiometric assay (IRMA) to detect intact PTH levels using different solid phase matrices. Different methods for immobilization of antibodies have also been evaluated. METHODS: Experiments were carried out with physical adsorption of antibodies, covalent coupling using 2 per cent glutaraldehyde and N,N`carbonyldiimidazole. In all cases, antibodies raised against C-terminal were used as solid phase agent. Detector antibodies were N terminal antibodies that were radio-iodinated with [125] I followed by gel purification. Several of the antibodies coupled to various solid phase matrices were incubated with PTH standards and the detector antibody as well as the commercially available tracer from DiaSorin kit to identify a suitable match pair. RESULTS: The best pair was polyclonal C-terminal PTH antibody along with the kit tracer from DiaSorin with regards to antibody coated to magnetic cellulose particles. Among the various antibodies and the solid phases evaluated, the best assay was obtained with the matched pair of antibodies (70×G67 and 70×G68) from Fitzgerald immobilized on polystyrene tubes. The polyclonal antibody against C-terminal PTH was chosen as the capture antibody and [125] I labelled polyclonal antibody against N-terminal PTH as the tracer. The sample values obtained in the antibody coated tubes were comparable to those obtained using a commercial kit. INTERPRETATION & CONCLUSIONS: The results indicated the feasibility of adopting this system for further development into a PTH IRMA for regular production as there is no indigenous kit available for intact PTH.


Subject(s)
Antibodies, Immobilized , Bone Diseases, Metabolic/diagnosis , Immunoradiometric Assay/methods , Immunoradiometric Assay/standards , Parathyroid Hormone/blood , Glutaral , Humans , Imidazoles , Iodine Radioisotopes , Sensitivity and Specificity
18.
J Assoc Physicians India ; 59: 698-704, 2011 Nov.
Article in English | MEDLINE | ID: mdl-22616335

ABSTRACT

INTRODUCTION: This study documents 25(OH)D status and bone mineral density (BMD) in women of reproductive (WR) and post-menopausal (PMW) age-groups in south India. SUBJECTS AND METHODS: Serum calcium (Ca), phosphorus (iP), albumin, alkaline phosphatase (ALP), creatinine, 25(OH)D and intact parathormone (N-tact PTH) of WR (n = 55) and PMW (n = 136) women were analyzed over a period of one year. Bone mineral Density (BMD) (Hologic, USA) was estimated using Caucasian data as reference. RESULTS: In both, WR and in PMW 25(OH)D deficiency (< 20 ng/ml), insufficiency (20-30 ng/ml) and replete states (> 30 ng/ml) were seen in 76%, 16.5%, 7.5% vs 70%, 23% and 7% respectively. PMW had lower BMD (gm/cm2) than WR at forearm ( P = < 0.001), hip trochanter (P = < 0.0001), lumbar spine antero-posterior (LSAP) (P = < 0.001) and lateral (LS Lateral) (P = < 0.001). Osteoporosis was seen at hip (15% and 28%), forearm (nil and 11%), LSAP (6% and 22%) and LS lateral (0% and 23%) among WR and PMW respectively. BMD did not correlate with any of the biochemical indices but correlated with BMD at other sites. CONCLUSIONS: Vitamin D deficiency coexists with low BMD in our study group. Serum 25(OH)D needs to be documented in women having low BMD. Calcium and vitamin D need to be supplemented as part of therapy in PMW.


Subject(s)
Bone Density , Postmenopause , Vitamin D Deficiency/epidemiology , Vitamin D/analogs & derivatives , Absorptiometry, Photon , Adult , Aged , Arm/diagnostic imaging , Biomarkers/blood , Calcium/blood , Cross-Sectional Studies , Female , Femur/diagnostic imaging , Humans , India/epidemiology , Lumbar Vertebrae/diagnostic imaging , Middle Aged , Parathyroid Hormone/blood , Prevalence , Vitamin D/blood , Young Adult
19.
Langmuir ; 26(14): 11699-704, 2010 Jul 20.
Article in English | MEDLINE | ID: mdl-20536164

ABSTRACT

Results are reported on an experimental study of the rheology of hydrate-forming water-in-oil emulsions. Density-matched concentrated emulsions were quenched by reducing the temperature and an irreversible transition was observed where the viscosity increased dramatically. The hydrate-forming emulsions have characteristic times for abrupt viscosity change dependent only on the temperature, reflecting the importance of the effect of subcooling. Mechanical transition of hydrate-free water-in-oil emulsions may require longer times and depends on the shear rate, occurring more rapidly at higher rates but with significant scatter which is characterized through a probabilistic analysis. This rate dependence together with dependence on subcooling reflects the importance of hydrodynamic forces to bring drops or particles together.

20.
Ayu ; 31(4): 410-6, 2010 Oct.
Article in English | MEDLINE | ID: mdl-22048531

ABSTRACT

Vatarakta is the major example of Vata vyadhi, caused due to avarana pathology. The scenario of Utthana Vatarakta occurred owing to the margavarana pathology, which can very well be correlated with atherosclerotic peripheral arterial disease. The literature enlists a number of Guggulu prayogas in the management of Vatarakta. An additional cavernous revise was indispensable to bring out the precise outcome of these products. Keeping these visions in mind, the particular comparative study was performed with Kaishora guggulu and Amrita guggulu, which are explained in the same context. This is a single-blind comparative clinical study with a pre-test and post-test design, wherein a minimum of 30 patients of either sex, suffering from Utthana Vatarakta, in an age limit of 16 to 70 years, were selected and randomly categorized into two groups. The 15 patients of group A were treated with oral administration of Tab Kaishora guggulu 1 g thrice a day and the group B patients with Tab Amrita guggulu of the same dose pattern with anupana of lukewarm water. The therapeutic effect of the treatment was assessed in both the groups based on specific subjective and objective parameters. The results obtained were analyzed statistically in both the groups and the comparative effect was assessed using the unpaired "t" -test. In the present study, 80% of the patients from both the groups had madhumeha (Diabetes mellitus), shonita mada (Hypertension) or both. Fifty percent of the patients in group A and nearly 60% of the patients in group B, suffering from Utthana Vatarakta, had the habit of smoking.In both the groups, a statistically significant improvement was observed in all the criteria of assessment. The outcome of the study revealed an identical therapeutic efficacy of Kaishora guggulu and Amrita guggulu in Utthana Vatarakta. The use of Kaishora guggulu or Amrita guggulu as shamana Aushadhas was a perfect selection in the management of rakta margavaranajanya Utthana Vatarakta.

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