MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

PAMELA measurements of cosmic-ray proton and helium spectra

Protons and helium nuclei are the most abundant components of the cosmic radiation. Precise measurements of their fluxes are needed to understand the acceleration and subsequent propagation of cosmic rays in our Galaxy. We report precision measurements of the proton and helium spectra in the rigidity range 1 gigavolt to 1.2 teravolts performed by the satellite-borne experiment PAMELA (payload for antimatter matter exploration and light-nuclei astrophysics). We find that the spectral shapes of these two species are different and cannot be described well by a single power law. These data challenge the current paradigm of cosmic-ray acceleration in supernova remnants followed by diffusive propagation in the Galaxy. More complex processes of acceleration and propagation of cosmic rays are required to explain the spectral structures observed in our data.     

Comparison of food selection and growth performance of koi carp, Cyprinus carpio L., and goldfish, Carassius auratus (L.) in mono- and polyculture rearing in tropical ponds

To compare the effect of polyculture against conventional monoculture on ornamental carp production, investigations on food selection and growth performance of koi carp (K), Cyprinus carpio L. and goldfish (G), Carassius auratus (L.) were conducted in a 11‐week rearing experiment in two monoculture (100% K and 100% G) and five polyculture (90% K–10% G, 70% K–30% G, 50% K–50% G, 30% K–70% G and 10% K–90% G) conditions in tropical ponds. There were three replicates for each treatment. Environmental conditions and food availability were similar in all the treatments. Ivlev's electivity index showed that both fish species avoided phytoplankton and preferred cladocerans to other zooplankton groups (copepods and rotifers) in monotypic conditions. However, in the polyculture treatments, the positive electivity of goldfish towards cladocerans reduced significantly (P<0.05), while the percentage of copepods, rotifers and phytoplankton in the gut content increased. No significant differences in weight gain, specific growth rate and deformities were recorded at harvest for koi carp between the different treatments (P>0.05). Even the survival rate of koi carp recorded above 90% in all the treatments. However, the goldfish recorded significantly better weight gain, specific growth rate and survival in monoculture (100% G), compared with the polyculture treatments (P<0.05). Goldfish deformities were lowest (P<0.05) in the monoculture treatment (2.42%). The number of marketable fish above a set size limit of 4 g total weight was significantly higher in the two monoculture treatments, compared with the five polyculture treatments (P<0.05). Keeping in view of the dietary similarities of koi carp and goldfish, and the aggressive nature of koi carp in polyculture, it is suggested to refrain from polyculture of goldfish and koi carp until further documentations relating to optimum stocking density and management of polyculture of ornamental carps are available.     

Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings

Nine isonitrogenous (35% crude protein approximately) and isocaloric (18.37 kJ g^?1) experimental diets (RLL20–BCFL40) were formulated with either raw or treated (inoculated with fish intestinal bacteria) Leucaena leucocephala leaf meal at 20%, 30% and 40% levels replacing other ingredients partially from a fish meal based reference diet (RD). Two specific strains of fish intestinal bacteria, Bacillus subtilis (isolated from Cyprinus carpio) and B. circulans (isolated from Oreochromis mossambicus) having extracellular cellulolytic and amylolytic activities, were used to inoculate Leucaena leaf meal for 15 days at 37°C. The crude fibre, cellulose and hemicellulose contents and the antinutritional factors, tannin, phytic acid and mimosine in the leaf meal decreased due to inoculation. However, free amino acids and fatty acids increased in the treated leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a RD. Both the inclusion level and type of Leucaena leaf meal in diets significantly affected the growth performance of rohu. Fish fed diets containing inoculated Leucaena leaf meal performed better in comparison with those with the RD. On the basis of growth response, feed conversion ratio, protein efficiency ratio and apparent net protein utilization, diet formulated with 30%Leucaena leaf meal inoculated with B. circulans resulted in the best performance of rohu fingerlings followed by diet with 40%B. subtilis inoculated Leucaena leaf meal. The apparent protein digestibility (APD) was better in fish fed diets containing B. circulans inoculated leaf meal. An increasing level of raw Leucaena leaf meal was associated with a decrease in the carcass protein content of rohu fingerlings. The activity of α‐amylase increased with the increasing level of treated leaf meal in diets. Cellulase activity increased with increasing level of inclusion of raw leaf meal, and was comparatively lower in fish fed diets with treated leaf meal. Activities of protease and lipase were higher in fish fed the RD. The results showed that it is possible to incorporate Leucaena leaf meal inoculated with enzyme‐producing fish intestinal bacteria in carp diets up to 40% level of inclusion.     

Überleben einerXanthomonas campestris pv.pelargonii-Mutante in einjähriger Komposterde

Zusammenfassung Das Überleben vonXanthomonas campestris pv.pelargonii wurde anhand einer Mutante in einjährigem Kompost untersucht. In der Versuchsvariante 1, bei der die Bakterien mit Komposterde vermischt wurden, konnte die Mutante Xc3b nach 15 Monaten nicht mehr nachgewiesen werden. Unter diesen Bedingungen war es den Bakterien möglich, 14 Monate im Kompost zu überleben. In der Versuchsvariante 2, bei der Komposterde mit Bakterien und zusätzlich noch mit infizierten Pelargonienstielen eingebracht wurden, konnten dieXanthomonas-Mutanten noch nach 26 Monaten nachgewiesen werden.

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Survival of a mutant ofXanthomonas campestris pv.pelargonii in one year old compost

Survival of phytopathogenicXanthomonas campestris pv.perlargonii in a one year old compost was investigated by use of a mutant. In the variant 1, in which the compost soil was mixed with the bacteria, the mutant Xc-3b could not be detected after 15 months. Under these conditions the bacteria were able to survive for 14 months in the compost. In the variant 2, in which the compost soil was mixed with bacteria andXanthomonas-infected geranium stems, it could be found even after 26 months.

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Mit 2 Abbildungen     

Effect of FMD vaccination on semen quality parameters in Karan Fries and Murrah buffalo bulls

Effect of Foot and Mouth disease (FMD) vaccination was studied on semen quality parameters of 19 Karan Fries (KF) and eight Murrah (MU) breeding bulls during the period 2002 to 2004 at Artificial Breeding Complex, NDRI, Karnal. A total of non-vaccinated 155 KF and 72 MU bulls' ejaculates were taken as control, while 169 KF and 51 MU bulls' ejaculates, collected after vaccination, were used to study the effect of vaccination stress. The results showed that FMD vaccination had no significant (P > 0.05) effect on ejaculate volume and total volume per day of semen in both KF and MU bulls. Volume of semen increased slightly during post-vaccination period in both the breeds. After FMD vaccination, there was significant (P < 0.01) decrease in mass activity (2.27 ± 0.06 vs. 1.67 ± 0.07 and 2.49 ± 0.09. vs. 1.75 ± 0.10, for KF and MU, respectively), initial motility (56.89 ± 0.03% vs. 44.62 ± 0.02% and 62.26 ± 0.04% vs. 47.08 ± 0.05%, for KF and MU, respectively), sperm concentration (754.19 ± 23.96 vs. 554.14 ± 22.95 × 10⁶/ml and 848.61 ± 33.65 vs. 571.57 ± 39.99 × 10⁶/ml, for KF and MU, respectively), and total sperm output per ejaculate (3,685.94 ± 158.40 vs. 2,781.54 ± 151.70 × 10⁶ and 2,218.75 ± 133.14 vs. 1,582.84 ± 158.20 × 10⁶, for KF and MU, respectively). Application of FMD vaccine had significantly (P < 0.05) adverse effect on most of the seminal attributes during post-vaccination in KF and MU buffalo bulls. So, the spermiograms affected following vaccination suggest that in bovines, the semen collection and preservation should be suspended till normal fertility of sperm is restored to avoid the failure of conception from artificial insemination using such semen.     

Prevalence,molecular fingerprinting and drug resistance profile of enterovirulent <Emphasis Type="Italic">Escherichia coli</Emphasis> isolates from free-ranging yaks of Tawang district,Arunachal Pradesh,India

Of 273 samples (rectal swab) collected from free-ranging yaks of Tawang district, Arunachal Pradesh, 42 Shiga toxin-producing Escherichia coli (STEC), six enteropathogenic E. coli (EPEC) and 27 enterotoxigenic E. coli (ETEC) strains were isolated. All the STEC and EPEC strains were further investigated for respective stx variants (for STEC only) and additional putative virulence factors. The 27 ETEC strains were also screened for characteristic enterotoxin gene(s) and colonization factors. Occurrence of ETEC was significantly (p < 0.05) higher in the diarrheic yaks and yaks of less than 1 year of age. Majority of enterovirulent E. coli isolates were resistant to amikacin, azithromycin, chloramphenicol, colistin, doxycycline, furazolidone, nalidixic acid, nitrofurantoin, streptomycin and tetracycline. Dendrogram, constructed with molecular fingerprinting profiles obtained from RAPD (Randomly Amplified Polymorphic DNA) and ERIC (Enterobacterial Repetitive Intergenic Consensus) PCR, placed the isolates in different clusters irrespective of their serotypes, virulence gene and drug resistance pattern. Collectively, the study indicates that yaks, being a potential reservoir of multidrug resistant STEC and EPEC, may represent significant risk to public health in this region. Higher recovery of ETEC isolates from yaks with diarrhea points out that ETEC may be a major determinant for repeated occurrence of diarrhea in yaks.     

Priority habitats for the conservation of large river fish in the Ganges river basin

• 1. Three classes of habitat used by groups of fish species classified as conservation and management priorities were developed for the Gerua River (also known as the Girwa River, Karnali River) in the Ganges river basin. This river is large (mean annual discharge ca 1500 m³ s^?1, up to 900 m wide), surrounded by protected lands of India and Nepal, and upstream of major diversions and river alterations.
• 2. Fish and habitat sampling was conducted at 45 sites from 2000 to 2003. Data were analysed for 2172 fish of 14 species. Species and life stages found occupying a statistically distinct subset of the river habitats were grouped to identify classes of river habitat for conservation.
• 3. Most species and life‐stage groups specialized on specific habitat conditions revealed by multivariate analyses of variance and a principal component analysis. The most numerous and diverse group (six species, 15 life stages) was associated with deep depositional habitats with sandy substrate. Two species covering three life stages were primarily oriented to erosional habitat marked by fast current velocity with pebble and cobble substrate. A third group of three species of adults and juveniles were intermediate in habitat use.
• 4. River conservation for fish faunas should maintain both erosional and depositional channel habitats with depths, substrates, and current velocity inclusive of the ranges reported. The erosional and depositional nature of the key habitats requires that rivers be maintained with flows capable of channel‐forming functions.

Copyright © 2006 John Wiley & Sons, Ltd.     

Extraction,purification and properties of soil hydrolases

Invertase, cellulase, phosphatases, protease and β-glucosidase were extracted from permanent pasture soil with 0.2 M phosphate buffer (pH 8) in the presence of 0.2 M EDTA. This extract was further treated with ammonium and salmine sulphates. Attempts were made to fractionate these enzyme activities by gel and anion-exchange chromatography. Specific activities were estimated in all fractions and some characteristics of the purified enzymes (optimum pH, temperature and substrate concentration, and K_m and V_max) were investigated. The results indicated that extracted enzyme activities occurred partly in soil as a carbohydrate-enzyme complex and partly as a humo-carbohydrate complex.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.