Genetic diversity and bottleneck analysis of Nagpuri buffalo breed of India based on microsatellite data

In this study, 25 heterologous bovine microsatellite markers have been used for the assessment of genetic diversity in Nagpuri buffalo, an important breed of Central India. For this, 48 DNA samples of unrelated individuals of Nagpuri buffalo were PCR amplified and microsatellite alleles were resolved in 6% denaturing, silver stained Urea-PAGE gel. Genotypic status of individuals at each locus was identified manually and data analysis carried out using POPGENE software. Observed number of alleles varied from 2 (ILSTS073 locus) to 8 (HEL13 & ILSTS058 loci) with a mean of 5.24 alleles per locus. Moderate level of heterozygosity (0.45) indicated sufficient genetic diversity existing in this buffalo population. PIC values for the microsatellite loci analysed, ranged from 0.10 (ILSTS0I9 locus) to 0.81 (ILSTS058 locus) with a mean of 0.53. No shift in the frequency distribution of alleles and a normal L-shaped curve indicated non-existence of any bottleneck in Nagpuri. The study thus highlights the usefulness of heterologous bovine microsatellite markers to assess the genetic variability in buffalo breds as well. Also various diversity indices suggest sufficient genetic variability within Nagpuri buffalo that can be utilized as initial guidelines for future breeding strategies and conservation. The article is published in the original.     

Isolation of 4-aminopyridine resistant mutants affecting alkali-insoluble glucan content of cell walls in Saccharomyces cerevisiae

Saccharomyces cerevisiae cells when grown on synthetic medium plates containing 10 mM of 4-aminopyridine (4-AP) undergo cell lysis. Using an ethylmethane sulfonate mutagenesis (EMS) screen, 4-AP resistant mutants (apr) were isolated which could grow on inhibitory concentration of 4-AP. Eighty mutants were obtained that were recessive, monogenic and formed two complementation groups. To identify genes, whose products might be interacting with the apr loci, extragenic suppressors were isolated, which reverted 4-AP resistance phenotype of apr mutants. The suppressors, when genetically characterized, were found to be recessive and represented two loci with overlapping functions. Representative alleles from apr mutants were analyzed for cell wall composition. They were found to have a higher amount of alkali-insoluble glucan signifying the role of alkali-insoluble glucan in cell wall maintenance.     

Penicillin acylases revisited: importance beyond their industrial utility

It is of great importance to study the physiological roles of enzymes in nature; however, in some cases, it is not easily apparent. Penicillin acylases are pharmaceutically important enzymes that cleave the acyl side chains of penicillins, thus paving the way for production of newer semi-synthetic antibiotics. They are classified according to the type of penicillin (G or V) that they preferentially hydrolyze. Penicillin acylases are also used in the resolution of racemic mixtures and peptide synthesis. However, it is rather unfortunate that the focus on the use of penicillin acylases for industrial applications has stolen the spotlight from the study of the importance of these enzymes in natural metabolism. The penicillin acylases, so far characterized from different organisms, show differences in their structural nature and substrate spectrum. These enzymes are also closely related to the bacterial signalling phenomenon, quorum sensing, as detailed in this review. This review details studies on biochemical and structural characteristics of recently discovered penicillin acylases. We also attempt to organize the available insights into the possible in vivo role of penicillin acylases and related enzymes and emphasize the need to refocus research efforts in this direction.     

Seasonal changes in blood biochemical and endocrine responses of different indigenous goat breeds of tropical island agro-ecological environment

The objective of the current study was to observe the impact of two seasons viz. summer (February–May) and monsoon (June–August) on the blood biochemical and hormonal responses in different indigenous goats of tropical island agro-ecological environment maintained under extensive management system. Sixty animals of three different indigenous goat breeds were included in the study: Andaman local goat (AL, n = 20), Andaman local?×?Malabari (AL?×?M, n = 20), and Teressa goat (n = 20). Sixty serum samples (n = 10/season/breed) from the three groups of animals were analyzed. Study revealed that there was a significant increase (p ≤ 0.05) in serum total protein in Teressa goats during summer than monsoon. Serum albumin showed significant variation (p ≤ 0.05) between AL and AL?×?M during summer whereas significant variation of albumin (p ≤ 0.05) was observed between AL and Teressa, AL?×?M and Teressa during monsoon season. Significant differences in serum albumin (p ≤ 0.05) were also observed in AL and AL?×?M during summer and monsoon seasons, respectively. The serum cortisol levels were significantly higher (p ≤ 0.05) in AL goats during summer than in monsoon season. Thus, the study could able to establish the seasonal variation in biochemical and hormonal values of indigenous goat breeds in hot and humid tropical island environment.     

Effectiveness of action in India to reduce exposure of Gyps vultures to the toxic veterinary drug diclofenac

Contamination of their carrion food supply with the non-steroidal anti-inflammatory drug diclofenac has caused rapid population declines across the Indian subcontinent of three species of Gyps vultures endemic to South Asia. The governments of India, Pakistan and Nepal took action in 2006 to prevent the veterinary use of diclofenac on domesticated livestock, the route by which contamination occurs. We analyse data from three surveys of the prevalence and concentration of diclofenac residues in carcasses of domesticated ungulates in India, carried out before and after the implementation of a ban on veterinary use. There was little change in the prevalence and concentration of diclofenac between a survey before the ban and one conducted soon after its implementation, with the percentage of carcasses containing diclofenac in these surveys estimated at 10.8 and 10.7%, respectively. However, both the prevalence and concentration of diclofenac had fallen markedly 7-31 months after the implementation of the ban, with the true prevalence in this third survey estimated at 6.5%. Modelling of the impact of this reduction in diclofenac on the expected rate of decline of the oriental white-backed vulture (Gyps bengalensis) in India indicates that the decline rate has decreased to 40% of the rate before the ban, but is still likely to be rapid (about 18% year(-1)). Hence, further efforts to remove diclofenac from vulture food are still needed if the future recovery or successful reintroduction of vultures is to be feasible.     

A Rotational Cylindrical fMRI Phantom for Image Quality Control

Purpose

A novel phantom for image quality testing for functional magnetic resonance imaging (fMRI) scans is described.

Methods

The cylindrical, rotatable, ~4.5L phantom, with eight wedge-shaped compartments, is used to simulate rest and activated states. The compartments contain NiCl₂ doped agar gel with alternating concentrations of agar (1.4%, 1.6%) to produce T₁ and T₂ values approximating brain grey matter. The Jacard index was used to compare the image distortions for echo planar imaging (EPI) and gradient recalled echo (GRE) scans. Contrast to noise ratio (_CNR) was compared across the imaging volume for GRE and EPI.

Results

The mean T₂ for the two agar concentrations were found to be 106.5±4.8, 94.5±4.7 ms, and T₁ of 1500±40 and 1485±30 ms, respectively. The Jacard index for GRE was generally found to be higher than for EPI (0.95 versus 0.8). The CNR varied from 20 to 50 across the slices and echo times used for EPI scans, and from 20 to 40 across the slices for the GRE scans. The phantom provided a reproducible CNR over 25 days.

Conclusions

The phantom provides a quantifiable signal change over a head-size imaging volume with EPI and GRE sequences, which was used for image quality assessment.     

Predictors of Duration and Treatment Failure of Severe Pneumonia in Hospitalized Young Nepalese Children

Background

Pneumonia in young children is still the most frequent cause of death in developing countries. We aimed to identify predictors for recovery and treatment failure in children hospitalized with severe pneumonia.

Methods

We enrolled 610 Nepalese children, aged 2 – 35 months from February 2006 to June 2008. Study participants were provided with standard treatment for pneumonia and followed up until discharge. Three multiple regression models representing clinical variables, clinical and radiological combined and all variables, including C-reactive protein (CRP) and viral etiology were used to assess the associations.

Results

The median age of study participants was 6 months with 493 (82%) infants and 367 (61%) males. The median time (IQR) till recovery was 49 (31, 87) hours and treatment failure was experienced by 209 (35%) of the children. Younger age, hypoxia on admission and radiographic pneumonia were independent predictors for both prolonged recovery and risk of treatment failure. While wasting and presence of any danger sign also predicted slower recovery, Parainfluenza type 1 isolated from the nasopharynx was associated with earlier resolution of illness. Gender, being breastfed, stunting, high fever, elevated CRP, presence of other viruses and supplementation with oral zinc did not show any significant association with these outcomes.

Conclusion

Age, hypoxia and consolidation on chest radiograph were significant predictors for time till recovery and treatment failure in children with severe pneumonia. While chest radiograph is not always needed, detection and treatment of hypoxia is a crucial step to guide the management of hospitalized children with pneumonia.     

Gastro-protective and Anti-stress Efficacies of Monomethyl Fumarate and a <Emphasis Type="Italic">Fumaria indica</Emphasis> Extract in Chronically Stressed Rats

Results of the very first experiments conducted to evaluate therapeutic potentials of a fumarate containing Fumaria indica extract and of fairly low daily oral doses of monomethyl fumarate for prevention of chronic unavoidable foot-shock stress-induced gastric ulcers, and possible involvement of diverse neuro-hormonal and oxidative process in their stress response desensitizing effects are reported and discussed in this article. Preventive effects of 21 daily oral 60, 120, and 240 mg/kg doses of a standardized 50 % methanolic F. indica extract (MFI) and 1.25, 2.50, and 5.00 mg/kg/day of pure monomethyl fumarate (MMF) were compared in rats subjected to one hour daily unavoidable foot-shocks. A pharmaceutically well-standardized Withania somnifera (WS) root extract was used as a reference herbal anti-stress agent in all experiments. Effects of the treatments on stress-induced alterations in body weight, adrenal and spleen weights, gastric ulcer and ulcer index, weight of glandular stomach, protective mucosal glycoprotein content, cellular proliferation, oxidative stress on stomach fundus, and brain tissues of male rats were quantified. Other parameters quantified were plasma corticosterone levels, brain monoamine levels, and expressions of the cytokines TNF-α, IL-10, and IL-1β in blood and brain of stressed and treated rats. Most but not every observed stress-induced anomalies were suppressed or completely prevented by both MFI and pure MMF treatments in dose-dependent manner. Qualitatively, the observed activity profiles of both of them were similar to those of WS dose tested. These results reveal that both MFI and MMF are potent gastro-protective agents against chronic unavoidable stress-induced ulcers and strongly suggest that they act as regulators or modulators of monoamine, corticosterone, and cytokine homeostasis.     

<Emphasis Type="BoldItalic">Dictyostelium discoideum</Emphasis> RabS and Rab2 colocalize with the Golgi and contractile vacuole system and regulate osmoregulation

Small-molecular-weight GTPase Rab2 has been shown to be a resident of pre-Golgi intermediates and is required for protein transport from the ER to the Golgi complex; however, Rab2 has yet to be characterized in Dictyostelium discoideum. DdRabS is a Dictyostelium Rab that is 80% homologous to DdRab1 which is required for protein transport between the ER and Golgi. Expression of GFP-tagged DdRab2 and DdRabS proteins showed localization to Golgi membranes and to the contractile vacuole system (CV) in Dictyostelium. Microscopic imaging indicates that the DdRab2 and DdRabS proteins localize at, and are essential for, the proper structure of Golgi membranes and the CV system. Dominant negative (DN) forms show fractionation of Golgi membranes, supporting their role in the structure and function of it. DdRab2 and DdRabS proteins, and their dominant negative and constitutively active (CA) forms, affect osmoregulation of the cells, possibly by the influx and discharge of fluids, which suggests a role in the function of the CV system. This is the first evidence of GTPases being localized to both Golgi membranes and the CV system in Dictyostelium.