N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yI)colcemid, a probe for different classes of colchincine-binding site on tubulin

The nature of binding of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-colcemid (NBD-colcemid), an environment-sensitive fluorescent analogue of colchicine, to tubulin was tested. This article reports the first fluorometric study where two types of binding site of colchincine analogue on tubulin were detected. Binding of NBD-colcemid to one of these sites equilibrates slsowly. NBD-colcemid competes with colchicine for this site. Binding of NBD-colcemid to this site also causes inhibition of tubulin self-assembly. In contrast, NBD-colcemid binding to the other site is characterised by rapid equilibration and lack of competition with colchicine. Nevertheless, binding to this site is highly specific for the cholchicine nucleus, as alkyl-NBD analogues have no significant binding activity. Fast-reaction-kinetic studies gave 1.76 × 105 M–1 s–1 for the association and 0.79 s–1 for the dissociation rate constants for the binding of NBD-colcemid to the fast site of tubulin. The association rate constants for the two phases of the slow site are 0.016 × 10–4 M–1 s–1 and 3.5 × 10–4 M–1 respectively. These two sites may be related to the two sites of colchicine reported earlier, with binding characteristics altered by the increased hydrophobic nature of NBD-colcemid.

Characterization of a 10- to 14-kilodalton protease-sensitive mycobacterium tuberculosis H37Ra antigen that stimulates human -yi T cells

gamma delta T-cell receptor-bearing T cells (gamma delta T cells) are readily activated by intracellular bacterial pathogens such as Mycobacterium tuberculosis. The bacterial antigens responsible for gamma delta T-cell activation remain poorly characterized. We have found that heat treatment of live M. tuberculosis bacilli released into the supernatant an antigen which stimulated human gamma delta T cells, gamma delta T-cell activation was measured by determining the increase in percentage of gamma delta T cells by flow cytometry in peripheral blood mononuclear cells stimulated with antigen and by proliferation of gamma delta T-cell lines with monocytes as antigen-presenting cells. Supernatant from heat-treated M. tuberculosis was fractionated by fast-performance liquid chromatography (FPLC) on a Superose 12 column. Maximal gamma delta T-cell activation was measured for a fraction of 10 to 14 kDa. Separation of the supernatant by preparative isoelectric focusing demonstrated peak activity at a pi of <4.0. On two-dimensional gel electrophoresis, the 10- to 14-kDa FPLC fraction contained at least seven distinct molecules, of which two had a pi of <4.5. Protease treatment reduced the bioactivity of the 10- to 14-kDa FPLC fraction for both resting and activated gamma delta T cells. Murine antibodies raised to the 10- to 14-kDa fraction reacted by enzyme-linked immunosorbent assay with antigens of 10 to 14 kDa in lysate of M. tuberculosis. In addition, gamma delta T cells proliferated in response to an antigen of 10 to 14 kDa present in M. tuberculosis lysate. gamma delta T-cell-stimulating antigen was not found in culture filtrate of M. tuberculosis but was associated,vith the bacterial pellet and lysate of M. tuberculosis. These results provide a preliminary characterization of a 10- to 14-kDa, cell-associated, heat-stable, low-pI protein antigen of M. tuberculosis which is a major stimulus for human gamma delta T cells.

Studies on the induced breeding and post-larval rearing of shing (Heteropneustes fossilis Bloch)

An experiment was conducted on induced breeding and fry rearing of shing, Heteropneustes fossilis (Bloch) in the Department of Aquaculture, Bangladesh Agricultural University for a period of four months from April to July 1994. Hatching rate was calculated at 21.50h and was found to be 45 to 55 % and the survival rate of larvae was 30 to 40 % at 26 to 29°C. Survival rate and growth rate of post larvae were found to be 50 to 60% and 96.6 to 117.2% respectively. Feed-3 (F3) showed the highest survival rate and growth rate of post larvae.

Histopathological studies of EUS affected shing, Heteropneustes fossilis (Bloch) from a fish farm of Mymensingh, Bangladesh

An investigation was carried out to observe histopathological changes in liver and kidney of suspected epizootic ulcerative syndrome (EUS)-affected shing fish, Heteropneustes fossilis (Bloch) collected from the "Agro-3 fish farm" situated at Boilor, Trishal, Mymensingh. Focal necrosis, haemorrhages and atrophy of the sinusoidal region were observed in the liver tissue. Fungal granulomas were found both in liver and kidney. In some cases fatty depositions were observed in all over the hepatic tissue. Degeneration and necrosis of renal tubular epithelial cells were also occurred. Missing of glomerulus and necrosis surrounding the Bowman's capsule in the kidney tissue were found.

Experimental pathogenesis of Aeromonas hydrophila bacteria in shing Heteropneustes fossilis(Bloch)

Pathogenicity of Aeromonas hydrophila bacteria was tested on the stinging catfish Heteropneustes fossilis. Before artificial infection the morphological, biochemical and physiological characters of Aeromonas hydrophila were studied. The infections were done by two different methods, viz., intramuscular (IM) and intraperitoneal (IP) injection. In infection experiment, each group of 10 fish were injected either intramuscularly or intra peritoneally with one dose higher than the LD50 dose (9.6 x 107 CFU/fish). All the fish tested died within 1 to 9 days. Both in cases of intramuscular and intraperitoneal injection, external pathology were found. Haemorrhagic lesions were evident at the site of injection. The posterior end of the body surface was found to develop greyish-white lesion that was extended up to caudal fin. Hyperemic anal region and the fin bases were also observed. Total bacterial loads in liver, kidney and intestine were determined. Aeromonas hydrophila could be isolated from liver, kidney and intestine of the experimentally infected fish. In case of intramuscular injection the highest and the lowest bacterial load was found to be 2.4 x 107 CFU/g of liver and 2.1 x 102 CFU/g of kidney and in case of intraperitoneal injection they were found to be 3.6 x 106 CFU/g of kidney and 1.2 x 104 CFU/g of kidney respectively. It was concluded that A. hydrophila could cause serious disease condition to Heteropneustes fossilis and its pathogenesis in the fish was also very efficient.

Effect of carp PG doses on induced breeding of Shing, Heteropneustes fossilis (Bloch)

Five hormone doses viz. 25, 50, 75, 100, and 125 mg of carp PG/kg of body weight of the recipient fish were tested and they were designated as T1 T2, T3, T4, and T5 respectively. Significantly higher fertilization (98%) and hatching rates (38%) were obtained from T3 (75 mg of carp PG extract/kg body weight). While T4 (100 mg of carp PG extract/kg body weight) and T5 (125 mg of carp PG extract/kg body weight) gave the highest (90%) ovulation rate. In June and July the highest fertilization rate of 96 and 96.4% respectively and hatching rate 42.5 and 48.7% respectively were obtained. In over all consideration carp PG extract at a dose of 75 mg/kg body weight appears to be the suitable dose for induced breeding of H. fossilis and June and July are the suitable time for its induced breeding.

Proteomic analysis of colorectal cancer metastasis:stathmin-1 revealed as a player in cancer cell migration and prognostic marker

Metastasis accounts largely for the high mortality rate of colorectal cancer (CRC) patients. In this study, we performed comparative proteome analysis of primary CRC cell lines HCT-116 and its metastatic derivative E1 using 2-D DIGE. We identified 74 differentially expressed proteins, many of which function in transcription, translation, angiogenesis signal transduction, or cytoskeletal remodeling pathways, which are indispensable cellular processes involved in the metastatic cascade. Among these proteins, stathmin-1 (STMN1) was found to be highly up-regulated in E1 as compared to HCT-116 and was thus selected for further functional studies. Our results showed that perturbations in STMN1 levels resulted in significant changes in cell migration, invasion, adhesion, and colony formation. We further showed that the differential expression of STMN1 correlated with the cells' metastatic potential in other paradigms of CRC models. Using immunohistochemistry, we also showed that STMN1 was highly expressed in colorectal primary tumors and metastatic tissues as compared to the adjacent normal colorectal tissues. Furthermore, we also showed via tissue microarray analyses of 324 CRC tissues and Kaplan-Meier survival plot that CRC patients with higher expression of STMN1 have poorer prognosis.

Le prince Yi Hou [i. e. Yi Un] et la princesse Masako après leur mariage : [photographie de presse] / [Agence Rol]

Référence bibliographique : Rol, 60269

Un groupe de demoiselles d'honneur au mariage du prince Yi Hou [i. e. Yi Un] et la princesse Masako : [photographie de presse] / [Agence Rol]

Référence bibliographique : Rol, 60268