Histoenzymological comparison of the prostate gland of sexually 'quiescent' and 'active' Taphozous melanopogon melanopogon Temmnick (Microchiroptera: Mammalia)

Lysosomal hydrolases (e.g., acid phosphatase, AcPase; adenosine triphosphatase, ATPase, and lipase) and the mitochondrial 'marker' enzyme succinic dehydrogenase (SDH) were evaluated histochemically in the prostate gland of sexually 'quiescent' and 'active' bats. During the former state, AcPase activity was significantly less than in sexually active animals, suggesting that prostate AcPase activity is androgen dependent. Levels of lipase activity also were highest in the prostate of sexually active bats, suggesting the importance of endogenous lipids which may be mobilized and used as a source of energy. SDH and ATPase sites and patterns of distribution in the prostate gland of bats were closely similar during the two reproductive states. Differential enzymological patterns do not seem to have any significant correlation with the morphological changes which occur in the glandular epithelium, musculature, urethra and the luminal fluid, as the animals pass from a 'quiescent' phase to one of activity and vice versa as observed in the present study.     

Rab11-FIP3 is critical for the structural integrity of the endosomal recycling compartment

Rab11-FIP3 is an endosomal recycling compartment (ERC) protein that is implicated in the process of membrane delivery from the ERC to sites of membrane insertion during cell division. Here we report that Rab11-FIP3 is critical for the structural integrity of the ERC during interphase. We demonstrate that knockdown of Rab11-FIP3 and expression of a mutant of Rab11-FIP3 that is Rab11-binding deficient cause loss of all ERC-marker protein staining from the pericentrosomal region of A431 cells. Furthermore, we find that fluorophore-labelled transferrin cannot access the pericentrosomal region of cells in which Rab11-FIP3 function has been perturbed. We find that this Rab11-FIP3 function appears to be specific because expression of the equivalent Rab11-binding deficient mutant of Rab-coupling protein does not perturb ERC morphology. In addition, we find that other organelles such as sorting and late endosomes are unaffected by loss of Rab11-FIP3 function. Finally, we demonstrate the presence of an extensive coiled-coil region between residues 463 and 692 of Rab11-FIP3, which exists as a dimer in solution and is critical to support its function on the ERC. Together, these data indicate that Rab11-FIP3 is necessary for the structural integrity of the pericentrosomal ERC.     

Genetic regulation of gene expression during shoot development in Arabidopsis

The genetic control of gene expression during shoot development in Arabidopsis thaliana was analyzed by combining quantitative trait loci (QTL) and microarray analysis. Using oligonucleotide array data from 30 recombinant inbred lines derived from a cross of Columbia and Landsberg erecta ecotypes, the Arabidopsis genome was scanned for marker-by-gene linkages or so-called expression QTL (eQTL). Single-feature polymorphisms (SFPs) associated with sequence disparities between ecotypes were purged from the data. SFPs may alter the hybridization efficiency between cDNAs from one ecotype with probes of another ecotype. In genome scans, five eQTL hot spots were found with significant marker-by-gene linkages. Two of the hot spots coincided with classical QTL conditioning shoot regeneration, suggesting that some of the heritable gene expression changes observed in this study are related to differences in shoot regeneration efficiency between ecotypes. Some of the most significant eQTL, particularly those at the shoot regeneration QTL sites, tended to show cis-chromosomal linkages in that the target genes were located at or near markers to which their expression was linked. However, many linkages of lesser significance showed expected "trans-effects," whereby a marker affects the expression of a target gene located elsewhere on the genome. Some of these eQTL were significantly linked to numerous genes throughout the genome, suggesting the occurrence of large groups of coregulated genes controlled by single markers.     

Infectious salmon anaemia virus (ISAV) isolated from the ISA disease outbreaks in Chile diverged from ISAV isolates from Norway around 1996 and was disseminated around 2005, based on surface glycoprotein gene sequences

Background

All viruses in the family Bunyaviridae possess a tripartite genome, consisting of a small, a medium, and a large RNA segment. Bunyaviruses therefore possess considerable evolutionary potential, attributable to both intramolecular changes and to genome segment reassortment. Hantaviruses (family Bunyaviridae, genus Hantavirus) are known to cause human hemorrhagic fever with renal syndrome or hantavirus pulmonary syndrome. The primary reservoir host of Sin Nombre virus is the deer mouse (Peromyscus maniculatus), which is widely distributed in North America. We investigated the prevalence of intramolecular changes and of genomic reassortment among Sin Nombre viruses detected in deer mice in three western states.

Methods

Portions of the Sin Nombre virus small (S) and medium (M) RNA segments were amplified by RT-PCR from kidney, lung, liver and spleen of seropositive peromyscine rodents, principally deer mice, collected in Colorado, New Mexico and Montana from 1995 to 2007. Both a 142 nucleotide (nt) amplicon of the M segment, encoding a portion of the G2 transmembrane glycoprotein, and a 751 nt amplicon of the S segment, encoding part of the nucleocapsid protein, were cloned and sequenced from 19 deer mice and from one brush mouse (P. boylii), S RNA but not M RNA from one deer mouse, and M RNA but not S RNA from another deer mouse.

Results

Two of 20 viruses were found to be reassortants. Within virus sequences from different rodents, the average rate of synonymous substitutions among all pair-wise comparisons (π_s) was 0.378 in the M segment and 0.312 in the S segment sequences. The replacement substitution rate (π_a) was 7.0 × 10^-4 in the M segment and 17.3 × 10^-4 in the S segment sequences. The low π_a relative to π_s suggests strong purifying selection and this was confirmed by a Fu and Li analysis. The absolute rate of molecular evolution of the M segment was 6.76 × 10^-3 substitutions/site/year. The absolute age of the M segment tree was estimated to be 37 years. In the S segment the rate of molecular evolution was 1.93 × 10^-3 substitutions/site/year and the absolute age of the tree was 106 years. Assuming that mice were infected with a single Sin Nombre virus genotype, phylogenetic analyses revealed that 10% (2/20) of viruses were reassortants, similar to the 14% (6/43) found in a previous report.

Conclusion

Age estimates from both segments suggest that Sin Nombre virus has evolved within the past 37–106 years. The rates of evolutionary changes reported here suggest that Sin Nombre virus M and S segment reassortment occurs frequently in nature.     

Estimation of ileal output of gastro-intestinal glycoprotein in weaned piglets using three different methods

Mucin is the main constituent of gastrointestinal mucus and is responsible for its physicochemical and physiological properties. Previous studies have suggested that this glycoprotein represents a major component of undigested endogenous protein at the ileum. The aim of the study was to estimate the ileal output of this glycoprotein using three methods: direct ELISA, hexosamine-based method and ethanol precipitation. For setting up the ELISA assay, the glycoprotein was isolated from intestinal mucus scraping by cesium chloride density gradient ultracentrifugation and a rabbit hyperimmune plasma was raised against the purified glycoprotein. Ileal outputs of hexosamine and glycoprotein were measured in weaned piglets fed a control diet (C) based on casein or diets which contained 50% crude protein supplied by white (WCP) or black (BCP) chickpea. The hexosamine output was higher (P < 0.05) with the WCP diet (2.3 and 1.5 g x kg(-1) of dry matter intake for glucosamine and galactosamine, respectively) than with diet C (1.1 and 0.7 g x kg(-1) of DMI). The hexosamine-based and ethanol precipitation methods, but not the ELISA, showed significant differences between the diet treatments (P < 0.05). Although hexosamine-based and ethanol precipitation methods for the estimation of ileal glycoprotein appeared to be more satisfactory than the developed ELISA to display diet effects in this study, it remains to be determined whether the higher glycoprotein data variability observed with ELISA reflects the actual biological variability of the phenomenon or not.     

Blue through UV polarization sensitivities in insects

A Signal-to-Noise optimization model has now been extended to explain the range of species-specific polarization sensitivities of insects. The different polarization sensitivities are shown to represent optimizations for the detection of plane polarized (Rayleigh-scattered) skylight over a range of atmospheric polarization conditions. Rhodopsin absorption spectra with peaks in the Blue (max450 nm) maximize detection efficiencies under conditions of high polarization. Rhodopsin absorption spectra peaking in the UV (max350 nm) maximize Signal-to-Noise Ratios for the detection of polarized skylight at the other extreme of low polarization.     

Methemoglobinemia in aluminium phosphide poisoning in rats

Aluminium phosphide (AlP) a grain fumigant is the leading cause of intentional poisoning in North India. The mechanisms involved in toxicity are not known and there is no antidote till date. The present study was carried out to investigate the oxygen free radical generation, methemoglobinemia and effect of methylene blue treatment on survival time in rat model of AlP poisoning. AlP (50 mg/kg, intragastric) was administered in one group and the other group received AlP + Methylene Blue (MB) (0.1%, 1 mg/kg/5 min, i.v.). Malonyldialdehyde (MDA) and methemoglobin (MeHb) levels were measured at 10 and 30 min intervals. Blood MDA levels increased at 10 and 30 min after AlP exposure with simultaneous rise in MeHb levels suggesting methemoglobinemia could be due to increased oxygen free radical generation. Methylene blue caused a significant fall in both the parameters with prolongation of survival time. It is concluded that AlP causes methemoglobinemia responding to methylene blue treatment.