Effect of probiotic supplementation on total lactobacilli, bifidobacteria and short chain fatty acids in 2-5-year-old children.

​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.

Health & nutritional status of HIV infected children in Hyderabad, India.

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.

Comparative Structural and Functional Analysis of Staphylococcus aureus Glucokinase with other Bacterial Glucokinases.

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.

Standardisation of a two-site PTH immunoradiometric assay using various solid phase formats.

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.

Vitamin D status and bone mineral density in women of reproductive and postmenopausal age groups: a cross-sectional study from south India.

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.

Vitamin D status in Andhra Pradesh : a population based study.

BACKGROUND & OBJECTIVE: Data on the vitamin D status of the population in a tropical country like India have seldom been documented. Vitamin D deficiency is presumed to be rare. We carried out this study to document the dietary habits, serum calcium, 25(OH)vitamin D [25(OH)D], and parathyroid hormone levels of urban and rural population in a State in south India. METHODS: A total of 943 healthy urban and 205 rural adult subjects and 76 urban and 70 rural healthy children were studied for their dietary pattern, serum calcium, phosphorus, alkaline phosphatase, 25(OH)D, and N-tact parathyroid hormone levels (N-tact PTH). RESULTS: The daily dietary calcium intake of both the urban and rural population was low compared to that of recommended daily/dietary allowances (RDA) issued by Indian Council of Medical Research (ICMR). Dietary calcium and phosphorus were significantly lower(P<0.0001) in both the rural adult and children compared to that of the urban adult and children. The dietary phytate to calcium ratio was significantly (P<0.0001) higher in rural adult and children compared to that of urban adult and children. N-tact PTH levels negatively correlated with 25(OH)D in rural (r=-0.24; P<0.002), in urban adult subjects (r=-0.12; P<0.0001) and in rural and urban children (r=-0.2; P<0.05). The 25(OH)D levels of rural adult subjects were significantly higher (P<0.001) than that of urban adult subjects in both males and female groups. The 25(OH)D levels of both the urban and rural children were low. INTERPRETATION & CONCLUSION: Low dietary calcium intake and 25(OH)D levels were associated with deleterious effect on bone mineral homeostasis. Prospective longitudinal studies are required to assess the effect on bone mineral density, a surrogate marker for fracture risk and fracture rates.

Protective effect ofBacopa monniera L. on cytarabine induced biochemical changes in chick embryo.

Bacopa monniera, a medicinal plant distributed throughout India. Cytosine arabinoside (1-ß-arabinofuranosylcytosine; Ara-C) is the most important antimetabolite chemotherapeutic drug used for acute leukemia. In this study we examined the chemoprotective property of an ethanolic extract of Bacopa monniera on biochemical changes in chick embryo. CA caused biochemical changes in a concentration and time dependent manner in amniotic fluid, liver and heart tissues. Ethanolic extract of BM given to chick embryo at doses of 2, 4, 6 mg per egg. There is significant decrease in biochemical levels of glucose, protein, urea, uric acid, creatinine and inorganic phosphorus. Enzymatic activities of alkaline phosphatase, lactate dehydrogenase, serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase and malatedehydrogenase were also decreased with dose dependent manner in amniotic fluid, liver and heart tissues.

Spectrophotometric determination of some phenols with sodium metaperiodate and aminophenols.

New spectrophotometric methods have been developed for the assay of some phenols by use of two pairs of reagents, m-aminophenol and periodate (for catechol, catecholamines, eugenol and guaiacol) and metol and periodate (for pyrogallol, gallic acid, propyl gallate, phloroglucinol and butylated hydroxyanisole). The methods are simple, sensitive, reproducible, accurate within +/- 1.0%, and applicable to the assay of antioxidants (gallic acid, propyl gallate and butylated hydroxyanisole) in oils and fats, catecholamines in dosage forms, and eugenol in clove oil.