Identification of the major tyrosine phosphorylated protein of capacitated hamster spermatozoa as a homologue of mammalian sperm a kinase anchoring protein.

The molecular basis of mammalian sperm capacitation is unique in that, it is associated with a protein kinase A (PKA) dependent upregulation of protein tyrosine phosphorylation. Therefore, PKA activity during capacitation would be crucial for the downstream events of protein tyrosine phosphorylation, and mechanisms may exist to ensure that PKA phosphorylates its specific substrate. This could be achieved by bringing PKA close to its substrate, a function normally carried out by an A-kinase anchoring protein (AKAP). We showed previously that cauda epididymidal spermatozoa of hamster undergo a capacitation-dependent increase in protein tyrosine phosphorylation. In the present study, evidence is provided that two major tyrosine phosphorylated proteins of molecular weight 97 and 83 kDa are the hamster homologues of mouse pro-AKAP82 and AKAP82, and have been designated as hamster pro-AKAP83 and AKAP83 respectively. Hamster AKAP83 resembled the mouse AKAP82 with respect to its molecular weight, pI (pH 5-5.5) and cDNA and amino acid sequences. Sequence analysis indicated that the primary structure of pro-AKAP83 was highly conserved and exhibited 91% identity with mouse and rat AKAP82. Further, the functional domains, namely the region involved in binding the regulatory subunit of PKA and the proteolytic cleavage site between pro-AKAP83 and AKAP83, were identical with that observed in rat and mouse pro-AKAP82 and AKAP82. Immunoblot analysis using polyclonal hamster anti-AKAP83 antibodies indicated that AKAP83 was present both in caput and cauda epididymidal spermatozoa. The antibody also identified the pro-AKAP82 and AKAP82 in mouse caput and cauda epididymidal spermatozoa. Immunofluorescence studies indicated that AKAP83 in hamster spermatozoa was localized along the length of principal piece of the tail. It was also demonstrated that hamster pro-AKAP83/AKAP83 gene expression was testis specific and was not expressed in other organs in either sex. This is the first report implicating AKAP in capacitation in rodents.     

Impact of Habitat Fragmentation on the Demography of Lion-tailed Macaque (<Emphasis Type="Italic">Macaca silenus</Emphasis>) Populations in the Rainforests of Anamalai Hills,Western Ghats,India

Habitat fragmentation is considered the most serious threat to primate conservation in the tropics, and understanding it effects on lion-tailed macaque is very important because most of the populations live in fragmented habitats. We examined demographic parameters of 9 lion-tailed macaque groups in 8 rain forest fragments with reference to fragment area, tree density, canopy cover, tree height, and total basal area of food trees. Group size ranged from 7 to 90 individuals but was not related to habitat variables. Birth and growth rates of groups did not differ significantly between small (n = 4) and large (n = 4) fragments. Tree density, canopy cover, and total basal of food trees all show strong positive correlations with fragment area. Growth rate correlates with tree density, but there are no other significant relationships between birth or growth rate and habitat variables. The percentage of immature individuals in the group is significantly positively associated with the total basal area of food trees, but not with any other habitat variable. Comparison of our data from this study with data available for the same population in 1996 indicates a slight decline in birth rate but an increase in total number of individuals, from 154 to 242. Of the 5 small fragment groups, 3 have increased in size since 1996 while the sizes of the other 2 groups have remained the same. Based on this study, we advocate that to manage the fragile lion-tailed macaque groups the following steps need to be taken: 1) create dispersal corridors between the fragments using fruit trees to facilitate male dispersal, 2) construct canopy bridges across the prevailing roads, 3) protect the fragments from further degradation, and 4) periodically monitor these populations for long-term conservation.     

Ribonucleolytic activity of bovine spermatozoal plasma and acrosomal membranes

We show here the presence of an integral, strong RNAase-A-like ribonucleolytic activity in the plasma and outer acrosomal membranes of bovine spermatozoa. A part of this activity is extractable by salt or detergent.     

Thermolabile ribonucleases from antarctic psychrotrophic bacteria: detection of the enzyme in various bacteria and purification from Pseudomonas fluorescens

Abstract Thirteen terrestrial psychrotrophic bacteria from Antarctica were screened for the presence of a thermolabile ribonuclease (RNAase-HL). The enzyme was detected in three isolates of Pseudomonas fluorescens and one isolate of Pseudomonas syringae . It was purified from one P. fluorescens isolate and the molecular mass of the enzyme as determined by SDS-PAGE was 16 kDa. RNAase-HL exhibited optimum activity around 40°C at pH 7.4. It could hydrolyse Escherichia coli RNA and the synthetic substrates poly(A), poly(C), poly(U) and poly(A-U). Unlike the crude RNAase from mesophilic P. fluorescens and pure bovine pancreatic RNAase A which were active even at 65°C, RNAase-HL was totally and irreversibly inactivated at 65°C.     

A novel sialyltransferase inhibitor AL10 suppresses invasion and metastasis of lung cancer cells by inhibiting integrin‐mediated signaling

Aberrant sialylation catalyzed by sialyltransferases (STs) is frequently found in cancer cells and is associated with increased cancer metastasis. However, ST inhibitors developed till now are not applicable for clinical use because of their poor cell permeability. In this study, a novel ST inhibitor AL10 derived from the lead compound lithocholic acid identified in our previous study is synthesized and the anti‐cancer effect of this compound is studied. AL10 is cell‐permeable and effectively attenuates total sialylation on cell surface. This inhibitor shows no cytotoxicity but inhibits adhesion, migration, actin polymerization and invasion of α‐2,3‐ST‐overexpressing A549 and CL1.5 human lung cells. Inhibition of adhesion and migration by AL10 is associated with reduced sialylation of various integrin molecules and attenuated activation of the integrin downstream signaling mediator focal adhesion kinase. More importantly, AL10 significantly suppresses experimental lung metastasis in vivo without affecting liver and kidney function of experimental animals as determined by serum biochemical assays. Taken together, AL10 is the first ST inhibitor, which exhibits potent anti‐metastatic activity in vivo and may be useful for clinical cancer treatment. J. Cell. Physiol. 223: 492–499, 2010. © 2010 Wiley‐Liss, Inc.     

Tolazoline antagonises ketamine–xylazine anaesthesia in an endangered Black buck (<Emphasis Type="Italic">Antilope cervicapra</Emphasis>)

Seventy-seven anaesthetic events were carried out in 22 captive adult Black bucks (Antilope cervicapra) of either sex with a combination of 2 mg kg⁻¹ ketamine hydrochloride with 0.25 mg kg⁻¹ xylazine hydrochloride using a dart delivered from a blowpipe. Randomised anaesthetised animals received an intravenous injection of either yohimbine hydrochloride (0.125 or 0.25 mg kg⁻¹) or tolazoline hydrochloride (1 or 2 mg kg⁻¹) after 30–40 min of anaesthesia to antagonise the anaesthetic effects. Ketamine–xylazine induced smooth, rapid and reliable anaesthesia within 5–7 min of darting with no clinical adverse effects and causalities during or post-anaesthesia. Yohimbine failed to antagonise the anaesthetic effects of ketamine–xylazine in the Black buck. On the other hand, tolazoline was found to be very effective in hastening recovery in dose-dependent manner within 0.5–1.5 min. This study documents the first report of ketamine–xylazine anaesthesia and its antagonism by tolazoline in captive Black buck.