Complete mitochondrial genome sequence of Catla catla and its phylogenetic consideration.

Complete nucleotide sequence of mitochondrial genome (mitogenome) of the Catla catla (Ostariophysi: Cypriniformes: Cyprinidae) was determined in the present study. Its length is 16,594 bp and contains 13 protein coding genes, 22 transfer RNAs, two ribosomal RNAs and one non-coding control region. Most of the genes were encoded on the H-strand, while the ND6 and eight tRNA (Gln, Ala, Asn, Cys, Tyr, Ser (UCN), Glu and Pro) genes were encoded on the L-strand. The reading frames of two pair of genes overlapped: ATPase 8 with 6 and ND4L with ND4 by seven nucleotides each. The main non-coding region was 929 bp, with three conserved sequence blocks (CSB-I, CSB-II, and CSB-III) and an unusual simple sequence repeat, (TA)(7). Phylogenetic analyses based on complete mitochondrial genome sequences were in favor of the traditional taxonomy of family Cyprinidae. In conclusion present mitogenome of Catla catla adds more information to our understanding of diversity and evolution of mitogenome in fishes.

Complete mitochondrial genome of Labeo rohita.

The complete mitochondrial genome of Labeo rohita, an important cultivable fish, was determined for the first time. The genome is 16,611 bp in length and consists of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and one control region. The gene organisation and its order were similar to other vertebrates. The overall base composition on heavy strand was as follows A: 32.5%, G: 15.2%, C: 27.7%, T: 24.47%, and the A+T content 56.9%. The control region contains a microsatellite, (TA)(12), a putative termination-associated sequence and three conserved sequence blocks. This mitogenome sequence data would play an important role in population genetics and phylogenetics of Indian major carps.

Study of AP endonuclease (APEX1/REF1), a DNA repair enzyme, in gallbladder carcinoma.

AIM: This study investigated the levels of Apurinic/Apyrimidinic Endonuclease (APEX1) in gallbladder carcinoma (CaGB) tissue and co-related these levels with various clinicopathological parameters. PATIENTS AND METHODS: Twenty cases of CaGB and cholelithiasis were included in the study. Western blot analysis of APEX1 protein was performed using actin as the reference point. Densitometric analysis and the integrated density value (IDV) of APEX1 protein samples were determined. The ratio of IDV of APEX1/actin was determined. RESULTS: The mean IDV ratio of APEX1 in CaGB was 0.63±0.33 and 0.45±0.19 in cholelithiasis. The mean IDV ratio of a variant of APEX1 (ΔAPEX1) in CaGB was 0.50±0.09, whereas it was 0.40±0.16 in cholelithiasis. Calculating the mean IDV ratio of total APEX (APEX1+ΔAPEX1) in CaGB was 1.13±0.31 whereas in cholelithiasis, 0.85±0.23. The differences were statistically significant (p<0.05). CONCLUSION: A significant correlation was found between the relative expressions of APEX1 in cancer as compared to that in cholelithiasis patients. There was significant association between APEX1 expression and perineural invasion. A variant of APEX1 correlated with tumor infiltration. Hence APEX1 may be of use as a prognostic marker in patients with CaGB.

Jejunal atresia associated with idiopathic ileal perforation.

Jejunoileal atresia is one of the common causes of neonatal intestinal obstruction. Intestinal perforation with meconium peritonitis in the neonatal period, which carries a high mortality rate, is also common. The association of jejunal atresia with idiopathic ileal perforation is very rare.

Effect of 5-fluorouracil and mitomycin on the healing of intestinal anastomosis.

OBJECTIVE: Antineoplastic agents affect the healing of intestinal anastomosis. The aim was to evaluate the effect of 5-fluorouracil (5-FU) and mitomycin on the healing of the intestinal anastomosis and their schedule of administration. MATERIAL AND METHODS: Eighty-nine male albino Charles Foster rats with a mean weight of 256.57 g were divided into six groups. Group A represents the control, while in others varying schedules of chemotherapy (5-FU and mitomycin) were administered. The sacrifices were made on days 7, 14 and 21 postoperatively and bursting pressure and hydroxyproline content were measured. RESULTS: Nine rats died before completion of the experiment and were excluded. Adhesions were noted in all rats on sacrifice. The mean bursting pressure of normal intestine (group A) was 252 mm Hg. The bursting pressure was lower on day 7 (208 mm Hg) and it subsequently increased by day 21 (230 mm Hg). The mean bursting pressure in groups B, C, D and E was 174, 194, 182 and 188 mm Hg and it subsequently increased to 232, 272, 244 and 286 mm Hg. There was no difference in the pattern of bursting pressure in colon and ileum. The mean hydroxyproline content of ileum (group A) on day 7 was 34.37 mg/g tissue. The hydroxyproline content of the ileum in groups B, C, D and E was 15.08, 27.03, 7.75 and 21.04 mg/g tissue. There was a significant decrease in hydroxyproline content following anastomosis and chemotherapy. CONCLUSIONS: The effect of chemotherapy was pronounced when administered on the day of surgery or in the immediate pre- or postoperative period. Hence administration of chemotherapy during this period may be harmful.

Nitrite toxicity in Indian major carps: sublethal effect on selected enzymes in fingerlings of Catla catla, Labeo rohita and Cirrhinus mrigala.

The effects of 96-h sublethal exposure of nitrite (1, 2, 4, 8 and 10.4 mg l(-1)) on selected enzymatic activities in serum and tissues of fingerlings of catla (Catla catla), rohu (Labeo rohita) and mrigal (Cirrhinus mrigala) were studied for the first time in these species. All three species responded almost identically to nitrite exposure. With increasing nitrite concentration, reduction in activities was observed in acetylcholinesterase (AChE) in brain and liver; alkaline phosphatase (ALP) in serum, brain and gill; and acid phosphatase (ACP) in gill, while progressive increase in alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) activities in brain, gill and serum, and ACP activity in serum and brain was observed. Lactate dehydrogenase (LDH) activity increased in gill, liver, kidney, brain and serum of all three species with increasing nitrite concentration up to 8 mg l(-1) followed by reduction at 10.4 mg l(-1). The study revealed nitrite stress causing alteration in activities of all measured tissue and serum enzymes in the fingerlings, and so stresses the need for proper management of this particular nutrient in water during carp culture.

Alteration of catecholamines in pheochromocytoma (PC12) cells in vitro by the metabolites of chlorotriazine herbicide.

The effects of four major chlorotriazine metabolites on the constitutive synthesis of the catecholamines dopamine (DA) and norepinephrine (NE) were examined, using undifferentiated PC12 cells. NE release and intracellular DA and NE concentrations were quantified, for up to 24 h after initiation of treatment with different concentrations, ranging from 0 to 400 microM, of the metabolites hydroxyatrazine (HA), 2-amino-4-chloro-6-isopropylamino-s-triazine (deethylchlorotriazine), 2-amino-4-chloro-6-ethylamino-s-triazine (deisopropylchlorotriazine), and diaminochlorotriazine (DACT). Hydroxyatrazine significantly decreased intracellular DA and NE concentrations in a dose- and time-dependent manner. This metabolite also caused a significant inhibition of NE release from the cells. In contrast, deethylchlorotriazine and deisopropylchlorotriazine significantly increased intracellular DA concentration following exposure to 50-200 microM from 12 to 24 h. Intracellular NE was significantly reduced at these same concentrations of deethylchlorotriazine at 24 h while the concentration of NE in PC12 cells exposed to deisopropylchlorotriazine was not altered at any dosage or time point measured. NE release was decreased at 18 (200 microM) and 24 h (100 and 200 microM) following exposure to deethylchlorotriazine and at 24 h (100 and 200 microM) following deisopropylchlorotriazine. DACT, at the highest concentration (160 microM), significantly increased intracellular DA and NE concentrations at 18 and 24 h. NE release was also increased at 40-160 microM at 24 h. The viability of the PC12 cells was tested using the trypan blue exclusion method. Following 18 to 24 h of treatments with HA, cell viability was reduced 10-12% at the two higher concentrations (200 and 400 microM), but, with other metabolites, the viability was reduced by only 2 to 5% at the highest concentrations. These data indicate that HA affects catecholamine synthesis and release in PC12 cells in a manner that is similar to synthesis of atrazine and simazine. On the other hand, deethylchlorotriazine and deisopropylchlorotriazine altered catecholamine synthesis in a manner similar to that observed in the rat brain following in vivo exposure (i.e., increased DA and decreased NE concentration), whereas DACT appeared to produce an increase in NE release as well as in the intracellular DA and NE concentrations. Overall, these findings suggest that the catecholamine neurons may be a target for the chlorotriazines and/or their metabolites, that the metabolites produce a unique pattern of catecholamine response, and that all of the changes were seen within the same range of doses.

Differential modulation of catecholamines by chlorotriazine herbicides in pheochromocytoma (PC12) cells in vitro.

Epidemiological, wildlife, and laboratory studies have pointed to the possible adverse health effects of chlorotriazine herbicide (i.e. , atrazine, simazine, and cyanazine) exposure. However, the cellular mechanism(s) of action of these compounds remains unknown. Recently, it was reported by Cooper et al. (2000, Toxicol. Sci. 53, 297-307) that atrazine disrupts ovarian function by altering hypothalamic catecholamine concentrations and subsequently the regulation of luteinizing hormone (LH) and prolactin (PRL) secretion by the pituitary. In this study, we examined the effect of three chlorotriazines on catecholamine metabolism in vitro using PC12 cells. Intracellular norepinephrine (NE) and dopamine (DA) concentrations and spontaneous NE release were measured following treatment with different concentrations of atrazine, simazine (0, 12. 5, 25, 50, 100, and 200 microM) and cyanazine (0, 25, 50, 100, and 400 microM) for 6, 12, 18, 24, and 48 h. Atrazine and simazine significantly decreased intracellular DA concentration in a concentration-dependent manner. Intracellular NE concentration was also significantly decreased by 100 and 200 microM atrazine and 200 microM simazine. Similarly, there was a dose-dependent inhibition of NE release with 100 and 200 microM concentrations of both compounds. Although 100 and 400 microM cyanazine increased intracellular NE concentration, 50, 100, and 400 microM cyanazine significantly increased NE release at 24 and 36 h. In contrast, intracellular DA concentration was decreased by cyanazine, but only at 400 microM. The GABA(A)-receptor agonist, muscimol (0, 0.01, 0.1, and 1.0 microM) had no effect on either the release or on intracellular catecholamine concentrations from 6 through 24 h of treatment. Cell viability was somewhat lower in the groups exposed to 100 and 200 microM atrazine and simazine. However, the reduction in viability was significant only in the highest dose of atrazine used (200 microM) at 24 h. Cyanazine did not have an effect on the viability at any of the doses tested, and the cells were functional, even up to 48 h of exposure. These data indicate that both atrazine and simazine inhibit the cellular synthesis of DA mediated by the tyrosine hydroxylase (TH), and NE mediated by dopamine beta-hydroxylase (DbetaH), and, as a result, there is a partial or significant inhibition of NE release. Cyanazine, on the other hand, stimulated the synthesis of intracellular NE, and not DA. Thus, chlorotriazine compounds presumably act at the enzymatic steps or sites of CA biosynthesis to modulate monoaminergic activity in PC12 cells.

Cleogynol, a novel dammarane triterpenoid from Cleome gynandra.

Chemical examination of Cleome gynandra (whole plant excluding seeds) led to the isolation and identification of a novel (20S, 24S)-epoxy-19,25-dihydroxydammarane-3-one hemiketal (1). The structure of the new compound, designated cleogynol, was determined using spectral and chemical methods.

Dried sera for confirming blood-borne virus infections (HCV, HTLV-I, HIV & HBsAg).

For safe blood transfusion, developing countries face considerable problems including serological screening and confirmation of blood-borne virus infections (HCV, HTLV-I, HIV and HBsAg). Confirmation tests are not only costly but also require sophisticated techniques and expertise. In order to provide this support we have attempted to perform a virus antibody confirmation test on samples dried on blotting paper (BP). Forty-nine sera derived from selected patients and donors from Bombay, and nine donors' sera from Bellarussia were transported on BP. In control experiments, dilutions of antibody-positive sera (HIV, HTLV-I & HCV) and 'blinded' HTLV-I antibody-positive and antibody-negative donors were applied on BP. Eluates from snipped BP were tested initially by screening tests, and the reactives were subjected to confirmatory tests for three types of virus antibody tests (HCV, HTLV-I & HIV) by blotting methods and neutralisation tests for HBsAg. There was considerable reduction of titres in dry sera but all BP-derived dry specimens gave excellent qualitative concordance with their liquid-equivalent sera, and the HTLV-I-positive donor was identified and reconfirmed correctly. Presence of only HCV antibody was confirmed in all the nine selected Bellarussian donors. Blood donors in Bombay had 3% HIV antibody, 6% HBsAg and none had HCV antibody, while selected patients showed substantially higher levels of these markers: HIV-antibody 64%, HBsAg 57% and HCV-antibody 17% confirmed positive. The cause of this high level remains to be established. Dry samples received by post seem to be an economical approach to a first step in providing some levels of independent confirmation of reactives in developing countries.

Rectal stricture: a complication of tuberculosis.

Tuberculosis of the rectum is rarely reported, even from areas where tuberculosis, and gastrointestinal tuberculosis in particular, is prevalent. The authors report a case of long tubercular stricture of the rectum and distal part of the sigmoid colon in a 12-year-old girl. Because of nonspecific symptoms and noncharacteristic radiological and endoscopic features, the diagnosis of this rare entity rests mainly on histological evidence of the classical tubercle in a surgical biopsy specimen.