Effect of 40-cm segment umbilical cord milking on hemoglobin and serum ferritin at 6 months of age in full-term infants of anemic and non-anemic mothers.

OBJECTIVE: To assess the effect of early clamping and milking of a 40-cm umbilical cord LUCM (long umbilical cord and milking) on hemoglobin (Hb) and serum ferritin concentrations at 6 months of age and to evaluate whether the effect is different in infants of anemic and non-anemic mothers. STUDY DESIGN: Eligible term-infants of anemic (maternal Hb<11.0 g dl(-1)) and non-anemic mothers (Hb ⩾11.0 g dl(-1)) were randomized to LUCM or control groups (N=100 each). In the LUCM group, the umbilical cord was clamped at 40-cm length and milked. The control group had the cord clamped at 5 cm and not milked. Neonatal morbidities until discharge and Hb and serum ferritin at 6 months of age were compared. Effects in infants of anemic and non-anemic mothers were compared. RESULT: Compared with infants of non-anemic mothers, cord Hb was similar (14.50±1.90 g dl(-1) vs 14.67±1.73 g dl(-1)), but cord ferritin lower (85.8±55.4 ng ml(-1) vs 119.4±58.5 ng ml(-1), P<0.01) in infants of anemic mothers. Mean Hb concentration at 6 months was 9.60±1.42 g dl(-1) in the LUCM group and 9.07±1.10 g dl(-1) in the control group (P=0.004). Mean serum-ferritin concentration at 6 months was 113.9±43.8 ng ml(-1) in the LUCM group and 70.8±39.5 ng/ml in the control group (P<0.001). The effectiveness of LUCM did not vary with the maternal anemia status. CONCLUSION: Keeping the umbilical cord long and milking may be an effective method for improving Hb and iron stores at 6 months of age in term-infants.

Hematobiochemical and pathological alterations due to chronic chlorpyrifos intoxication in indigenous chicken.

OBJECTIVE: The present study investigates the effect of oral administration of chlorpyrifos (CPF) in indigenous chicken. MATERIALS AND METHODS: The birds were divided into two groups I and II. Group I served as control and group II was treated with CPF (0.36 mg/kg) orally daily up to 12 weeks. Blood samples were assayed for hemoglobin (Hb), total erythrocyte count (TEC), total leukocyte count (TLC), differential leukocyte count, and biochemical constituents like alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholinesterase (CHE), total protein and uric acid. Representative pieces of tissues from liver and kidney were collected weekly for histopathological examination. RESULTS: A significant (P < 0.01) increase of Hb, TEC, TLC, and heterophil percent and decrease of lymphocyte percent was observed. Serum ALP, AST, ALT, and uric acid increased significantly and CHE values decreased significantly in CPF treated birds. The protein level remained similar. Uric acid level was found to be increased significantly in the treated group. The results indicate that chronic CPF intoxication produces hematological, biochemical, and pathological changes in treated birds.

Evaluation of Water Quality of River Narmada and Its Correlation with Occurrence of Enteropathogens.

Water quality of river Narmada was investigated employing the chemometric techniques and statistical analysis with a view to extract information about the variables responsible for spatial and temporal variation in physicochemical parameters. Analysis of Pearson's correlation revealed significant correlation among the physicochemical parameters while weak correlation with occurrence of enteropathogens. The multivariate statistics and principal component analysis applied to datasets indicated three to four components influencing the water to the extent of 78.172% (2003), 82.726% (2004), 83.819% (2005), 86.294% (2006), 85.952% (2007) and 76.620% (2008) of total variance. The water samples collected on seasonal basis during 2003-2008 revealed the presence of multidrug resistance enteric pathogens viz. Aeromonas, Enterobacter, Klebsiella, Salmonella, Serratia, Shigella and Vibrio throughout the study period. Thus seasonal effects, agricultural wastes, domestic and industrial waste water discharges and their organic load caused main variation in water quality of river Narmada. These results will assist in the water management of river water for varied future demands including human consumption, irrigation, industrial and river conservation.

Effect of concentration and addition method of glycerol on the quality of cryopreserved mithun (Bos frontalis) spermatozoa.

The effect of concentration and addition method of glycerol on the quality of cryopreserved mithun (Bos frontalis) spermatozoa was investigated. Semen samples were collected from five healthy mithun bulls through rectal massage method and cryopreserved in liquid nitrogen. The samples were diluted in Tris-egg yolk-glycerol extender, equilibrated for 4 h at 4 °C and loaded into 0.50-ml straws. The straws were then frozen in liquid nitrogen vapour for 10 min and finally plunged into liquid nitrogen for storage. The required amount of glycerol was added into the diluted samples either in a single dose (3%, 4%, 5%, 6% or 7%; added at 37 °C immediately before equilibration) or in split doses (5%, 6% or 7%; the total amount was divided into four equal parts, and a part was added at 37 °C immediately before equilibration, and the remaining parts were added subsequently at 1, 2 and 3 h of equilibration at 4 °C). In the single-dose addition method, following freeze-thawing, greater (p < 0.05) motility (%) and proportion of live spermatozoa with intact acrosome (LSIA, %) in 5% glycerol (40.6 ± 1.7 and 43.4 ± 1.8 respectively) and lesser (p < 0.05) total morphological abnormalities (%) in 5% (14.1 ± 0.8) and 6% (13.7 ± 1.0) glycerol were observed compared to the other glycerol concentrations. In the split-dose addition method, following freeze-thawing, greater (p < 0.05) motility (%) and LSIA proportion (%) were found in 5% (50.2 ± 1.9 and 53.3 ± 1.8 respectively) compared to 6% or 7% glycerol, but the total morphological abnormalities were not different among the glycerol concentrations. In addition, in all the glycerol concentrations, better (p < 0.05) post-freeze-thaw motility and LSIA proportions were observed when glycerol was added in split doses compared to a single dose. In conclusion, Tris-egg yolk extender with 5% glycerol added in split doses was found most suitable for cryopreserving mithun sperm.

Prediction of feed intolerance and necrotizing enterocolitis in neonates with absent end diastolic flow in umbilical artery and the correlation of feed intolerance with postnatal superior mesenteric artery flow.

OBJECTIVES: To evaluate the role of postnatal superior mesenteric artery (SMA) flow in predicting feed intolerance and NEC in the babies who had AEDF in comparison with gestation matched SGA and AGA with normal flow. DESIGN: This was a prospective cohort study conducted in 62 eligible babies admitted in NICU. Babies were enrolled in 3 groups. Group 1 (n = 23) was SGA and AEDF, group 2 (n = 20) was SGA and group 3 (n = 19) was AGA and both with normal UA flow. In all babies baseline SMA flow was measured before test feed (0.5 ml) and repeated every 15 minutes for 1 hour after the feed. RESULTS: Feed intolerance was seen in 69.5% of babies in group1 (p = <0.001) as compared to 20% and 17.5% in group 2 and 3. Four (17.3%) babies developed NEC in group1 (p = 0.02) but none in other 2 groups. Baseline peak systolic velocity (PSV) and time average mean velocity (TAMV) at 60 min post feed were significantly (p = 0.01 and 0.028 respectively) lower in group1 than group3. TAMV and PSV at 60 min post feed were significantly lower (p = 0.028 and 0.03) in babies with feed intolerance as compared to no feed intolerance group. Absent end diastolic flow and hypoglycemia were independent risk factors for feed intolerance. CONCLUSION: SGA babies with AEDF had higher incidence of feed intolerance and NEC. Serial SMA flow studies specially the 60 min post feed study may help in differentiating which babies are likely to develop feed intolerance.

Wound healing activity of methanolic extract of leaves of Alternanthera brasiliana Kuntz using in vivo and in vitro model.

Wound healing activity of methanolic extract of leaves of Alternanthera brasiliana Kuntz was studied by excision and incision wound model (in vivo) in Sprague Dawley rats and by Chorioallantoic membrane (CAM) model (In vitro) in 9-day-old embryonated chicken eggs. In excision wound model, compared to the control group, per cent contraction of wound was significantly higher in A. brasiliana (5% w/w ointment) treated group. In incision wound model, tensile strength of the healing tissue after treatment with A. brasiliana was found to be significantly higher compared to the control group indicating better wound healing activity of the test plant. These findings were also confirmed by histopathological examination. The extract also promoted angiogenesis as evidenced by CAM model. The results suggested that methanolic extract of A. brasiliana possess significant wound healing potential in normal wound.

Increase in weight induced by muraglitazar, a dual PPARalpha/gamma agonist, in db/db mice: adipogenesis/or oedema?

BACKGROUND AND PURPOSE: Muraglitazar, a dual PPARalpha/gamma agonist, caused a robust increase in body weight in db/db mice. The purpose of the study was to see if this increase in weight was due to oedema and/or adipogenesis. EXPERIMENTAL APPROACH: The affinity of muraglitazar at PPARalpha/gamma receptors was characterized using transactivation assays. Pre-adipocyte differentiation, expression of genes for adipogenesis (aP2), fatty acid oxidation (ACO) and sodium reabsorption (ENaCgamma and Na+, K+-ATPase); haemodilution parameters and serum electrolytes were measured to delineate the role of muraglitazar in causing weight gain vis a vis rosiglitazone. KEY RESULTS: Treatment with muraglitazar (10 mg kg(-1)) for 14 days significantly reduced plasma glucose and triglycerides. Reduction in plasma glucose was significantly greater than after similar treatment with rosiglitazone (10 mg kg(-1)). A marked increase in weight was also observed with muraglitazar that was significantly greater than with rosiglitazone. Muraglitazar increased aP2 mRNA and caused adipocyte differentiation in 3T3-L1 cells similar to rosiglitazone. It also caused a marked increase in ACO mRNA in the liver of the treated mice. Expression of mRNA for ENaCgamma and Na+, K+-ATPase in kidneys was up-regulated after either treatment. Increased serum electrolytes and decreased RBC count, haemoglobin and haematocrit were observed with both muraglitazar and rosiglitazone. CONCLUSIONS AND IMPLICATIONS: Although muraglitazar has a better glucose lowering profile, it also has a greater potential for weight gain than rosiglitazone. In conclusion, muraglitazar causes both robust adipogenesis and oedema in a 14-day treatment of db/db mice as observed in humans.

cDNA cloning of putative rat acetyl-CoA transporter and its expression pattern in brain.

Rat acetyl-CoA transporter gene (Acatn) encodes a hydrophobic multi-transmembrane protein involved in the O-acetylation of gangliosides. O-acetylated gangliosides have been found to play important roles in the embryonic development of the nervous system. We have isolated rat Acatn cDNA by PCR cloning. The amino acid sequence of rat Acatn exhibited 92% and 96% homology with human and mouse sequences, respectively. The mRNA was expressed in brain at all developmental stages. Acatn expression was higher in embryonic and postnatal rats than in adult rats. Cellular localization of Acatn mRNA in adult rat brain was also analyzed by in situ hybridization. Acatn mRNA expression was detected in the neuronal cells of cerebellum, hippocampus, hypothalamus, cortex, olfactory bulb, and dorsal and ventral anterior olfactory nucleus in adult rat brain.

Genomic structure and promoter analysis of putative mouse acetyl-CoA transporter gene.

The acetyl-CoA transporter gene (Acatn) encodes a hydrophobic, multitransmembrane protein that is involved in the process of O-acetylation of sialic acid residues on gangliosides. O-Acetylated gangliosides have been found to play important roles in tissue development and organization during early embryonic stages. We have cloned the gene for mouse acetyl-CoA transporter. The gene spans approximately 20 kb and is composed of seven exons and six introns. A single transcription initiation site, 371 bp upstream of the ATG start codon, was identified. The promoter region was found to lack a TATA box. However, several potential transcription factor binding motifs such as AP1, AP2, C/EBPalpha, C/EBPbeta, HSF, GATA2 and MZF1 were identified in the promoter region.

Cloning and characterization of a putative mouse acetyl-CoA transporter cDNA.

A mouse acetyl-CoA transporter (Acatn) cDNA was isolated by PCR cloning. Mouse Acatn exhibited 92% homology with human sequence on the basis of amino-acid sequence. The predicted gene product of Acatn is a 61 kDa hydrophobic protein with six to 10 transmembrane domains. Transfection of mouse Acatn cDNA into HeLa/GT3+ cells resulted in significant increase in the amount of 9-O-acetylated gangliosides, suggesting that Acatn does play an important role in the acetylation of gangliosides. Northern blot analysis of Acatn mRNA suggested that transcript of Acatn is widely distributed in various adult tissues. Expression of Acatn was found to be developmentally regulated, with high expression levels during early embryonic stages, and then there was a subsequent decrease in expression levels in the later embryonic stages.

Novel gene delivery to liver cells using engineered virosomes.

We have demonstrated for the first time that the reconstituted Sendai viral envelopes containing only the fusion protein (F-virosomes) are efficient vehicles for the delivery of foreign genes specifically into human hepatoblastoma cells (HepG2) in culture. The membrane fusion-mediated entry of CAT (chloramphenicol acetyl transferase) gene into the cells was confirmed and the amount delivered to various subcellular fractions was quantitated. The dose dependence and kinetics of expression of biologically active CAT protein in HepG2 cells was measured. The CAT expression level in F-virosome-mediated delivery was significantly higher than that of Lipofectin or liganded proteo-liposome-mediated gene transfer. This kind of targeted delivery by means of membrane fusion induced by viral envelope glycoprotein may have wide applications to various gene transfer strategies both in vitro and in vivo.

Evaluation of canines for accelerant detection at fire scenes.

In recent years, canines have been successfully used in fire investigations to detect accelerant residues. We set out to determine the lower limits at which canines could reliably detect potential accelerants. Measured amounts ranging from 10 to as little as 0.01 microL of gasoline, kerosene, and isopars were applied to preselected spots along a continuous sample path (25 to 40 feet long) made out of burned and unburned wood or nylon carpeting strips at the testing site. Two canines were led past this sample path at least three times and positive alerts and negative responses were recorded. Both dogs were generally able to alert on spots containing 0.01 microL or more of all three accelerants, at or beyond the purge and trap recovery and gas chromatographic detection method employed. The canines did alert occasionally on background, especially that containing traces of styrene residues, either purposely added in specific amounts or formed upon partial pyrolysis of carpeting material. The dogs alerted on sites containing 0.1 to 1.0 microL of freshly applied gasoline or kerosene placed at actual heavily damaged fire scenes, but were less successful on samples containing smaller amounts.

Introduction of a Lepidopteran-Specific Insecticidal Crystal Protein Gene of Bacillus thuringiensis subsp. kurstaki by Conjugal Transfer into a Bacillus megaterium Strain That Persists in the Cotton Phyllosphere.

A lepidopteran toxin gene of the entomopathogen Bacillus thuringiensis subsp. kurstaki HD-1 was introduced into a cotton leaf-colonizing Bacillus megaterium strain, RS1, by conjugal transfer. Rifampin- and nalidixic acid-resistant colonies obtained after cell mating were screened for crystal production by microscopy. A transcipient, B. megaterium RS1-43, was selected by this procedure. Southern blot hybridization with both total DNA and HindIII-digested DNA of the transcipient showed positive signals with a cryIA-specific probe, suggesting the transfer of the lepidopteran-specific cryIA(a) gene. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis confirmed the presence of the 134-kDa toxic crystal protein specific to lepidopteran larvae in the transcipient. Survival studies with cultures of the transcipient at both vegetative and postvegetative growth stages on cotton, under field conditions, suggested that the bacterium persisted on the leaf surfaces for more than 28 days, with a gradual decline in the population level with time, while the donor, B. thuringiensis subsp. kurstaki, disappeared completely after 7 days following inoculation. An in situ differential crystal-staining technique indicated the production of crystals by the transcipient on cotton leaf surfaces for about 30 days. Leaf bioassays of cotton plants inoculated with a single spray of the transcipient showed 75- to 96% mortality to the first-instar larvae of Heliothis armigera up to 21 days, and this single spray conferred total protection to the plants for about 30 days by causing an antifeeding effect on the remaining larvae.