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.

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.

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.