Sand fly (Diptera: Psychodidae) distribution in the endemic and non‐endemic foci of visceral leishmaniasis in northwestern Iran

An entomological study was conducted from June to September, 2010 in rural regions of Azarbayjan‐e‐sharqi, Azarbayjan‐e‐qarbi, and Ardabil provinces in northwestern Iran to determine sand fly fauna, diversity, and distribution in different habitats and altitudes using both sticky papers and light traps. Geographical distribution of sand flies and the similarity of populations in different locations were analyzed ecologically based on the Shannon‐Wiener Index and Jacard Coefficient, respectively. A total of 3,982 specimens was collected and sixteen species recorded. They belonged to the genera Phlebotomus [subgenus Phlebotomus (P. papatasi), Paraphlebotomus (P. sergenti, P. mongolensis, P. caucasicus, P. jacusieli), Larroussius (P. major s.l., P. tobbi, P. perfiliewi transcaucasicus, P. kandelakii) and Adlerius (P. halepensis, P. brevis, P. longiductus, P. balcanicus)], and Sergentomyia [subgenus Sergentomyia (S. sintoni, S. dentata and S. theodori)]. P. papatasi was the predominant species in all the locations except Bileh Savar, Macu, and Meshkin Shahr, followed by P. perfiliewi transcaucasicus and P. kandelakii. The latter species were caught from different habitats and altitudes with higher frequency than other species of the subgenus Larroussius. The lowest abundance belonged to P. jacusieli. The predominant species of subgenus Adlerius was P. halepensis. Data analysis showed that Meshkin Shahr and Bileh Savar had high and low diversities of sand fly distribution, respectively. Meshkin Shahr and Sarab districts had the highest similarity. Both are located in the foothills of Sabalan Mountain, with high diversity and richness.     

Reduced efficacy of multiple doses of CpG-matured dendritic cell tumor vaccine in an experimental model

CpG motifs have been advanced as agents that stimulate the maturation of DCs for immunotherapy. The present study tested the hypothesis that multiple doses of CpG-matured DC vaccine would be efficacious for complete eradication of experimentally-induced tumor. Accordingly, WEHI164 cells were implanted subcutaneously in the flank of BALB/c mice. During DC culture, tumor lysate was added to immature DCs followed by addition of CpG or non-CpG control 4–6 h later. A total of three doses of CpG or non-CpG control-matured DCs were injected around tumors. The results showed that multiple doses of CpG-matured DCs led to considerable decrease in cytotoxicity of lymphocytes and significantly increased tumor growth rate compared to a single dose. Further, mice which received three doses of the vaccine also displayed significant FoxP3 in tumor tissue. In conclusion, multiple doses of CpG-matured DCs exhibited decreased anti-tumor immunity in association with increased expression of FoxP3.     

Glutathione-S-transferase P1 is a critical regulator of Cdk5 kinase activity

Cyclin dependent kinase-5 (Cdk5) activity is deregulated in Alzheimer's disease (AD) and contributes to all three hallmarks: neurotoxic β-amyloid formation, neurofibrillary tangles, and neuronal death. However, the mechanism leading to Cdk5 deregulation remains controversial. Cdk5 deregulation in AD is usually linked to the formation of p25, a proteolysis product of Cdk5 activator p35, which leads to Cdk5 mislocalization and hyperactivation. A few studies have indeed shown increased p25 levels in AD brains; however, others have refuted this observation. These contradictory findings suggest that additional factors contribute to Cdk5 deregulation. This study identified glutathione-S-transferase pi 1 (GSTP1) as a novel Cdk5 regulatory protein. We demonstrate that it is a critical determinant of Cdk5 activity in human AD brains and various cancer and neuronal cells. Increased GSTP1 levels were consistently associated with reduced Cdk5 activity. GSTP1 directly inhibits Cdk5 by dislodging p25/p35, and indirectly by eliminating oxidative stress. Cdk5 promotes and is activated by oxidative stress, thereby engaging a feedback loop which ultimately leads to cell death. Not surprisingly, GSTP1 transduction conferred a high degree of neuroprotection under neurotoxic conditions. Given the critical role of oxidative stress in AD pathogenesis, an increase in GSTP1 level may be an alternative way to modulate Cdk5 signaling, eliminate oxidative stress, and prevent neurodegeneration.     

Genomic,Proteomic, Morphological,and Phylogenetic Analyses of vB_EcoP_SU10, a Podoviridae Phage with C3 Morphology

A recently isolated phage, vB_EcoP_SU10 (SU10), with the unusual elongated C3 morphotype, can infect a wide range of Escherichia coli strains. We have sequenced the genome of this phage and characterized it further by mass spectrometry based proteomics, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and ultra-thin section electron microscopy. The genome size is 77,327 base pairs and its genes, and genome architecture, show high similarity to the phiEco32 phage genes and genome. The TEM images reveal that SU10 have a quite long tail for being a Podoviridae phage, and that the tail also changes conformation upon infection. The ultra-thin section electron microscopy images of phages at the stage of replication within the host cell show that the phages form a honeycomb-like structure under packaging of genomes and assembly of mature capsids. This implies a tight link between the replication and cutting of the concatemeric genome, genome packaging, and capsid assembly. We have also performed a phylogenetic analysis of the structural genes common between Podoviridae phages of the C1 and C3 morphotypes. The result shows that the structural genes have coevolved, and that they form two distinct groups linked to their morphotypes. The structural genes of C1 and C3 phages appear to have diverged around 280 million years ago applying a molecular clock calibrated according to the presumed split between the Escherichia – Salmonella genera.     

Three phases hollow fiber LPME combined with HPLC-UV for extraction, preconcentration and determination of valerenic acid in Valeriana officinalis

In the present work, the applicability of hollow fiber-based liquid phase microextraction (HF-LPME) was evaluated for the extraction and preconcentration of valerenic acid prior to its determination by reversed-phase HPLC/UV. The target drug was extracted from 5.0 mL of aqueous solution with pH 3.5 into an organic extracting solvent (dihexyl ether) impregnated in the pores of a hollow fiber and finally back extracted into 10 μ L of aqueous solution with pH 9.5 located inside the lumen of the hollow fiber. In order to obtain high extraction efficiency, the parameters affecting the HF-LPME, including pH of the donor and acceptor phases, type of organic phase, ionic strength, the volume ratio of donor to acceptor phase, stirring rate and extraction time were studied and optimized. Under the optimized conditions, enrichment factor up to 446 was achieved and the relative standard deviation (RSD) of the method was 4.36% (n = 9). The linear range was 7.5-850 μg L?1 with correlation coefficient (r2=0.999), detection limits was 2.5 μg L?1 and the LOQ was 7.5 μg L?1. The proposed method was evaluated by extraction and determination of valerenic acid in some Iranian wild species of Valerianaceae.     

Joint analysis of beef growth and carcass quality traits through calculation of co-variance components and correlations

A joint growth-carcass model using random regression was used to estimate the (co)variance components of beef cattle body weights and carcass quality traits and correlations between them. During a four-year period (1994-1997) of the Australian "southern crossbreeding project", mature Hereford cows (N = 581) were mated to 97 sires of Jersey, Wagyu, Angus, Hereford, South Devon, Limousin, and Belgian Blue breeds, resulting in 1141 calves. Data included 13 (for steers) and 8 (for heifers) body weight measurements approximately every 50 days from birth until slaughter and four carcass quality traits: hot standard carcass weight, rump fat depth, rib eye muscle area, and intramuscular fat content. The mixed model included fixed effects of sex, sire breed, age (linear, quadratic and cubic), and their interactions between sex and sire breed with age. Random effects were sire, dam, management (birth location, year, post-weaning groups), and permanent environmental effects, and their interactions with linear, quadratic and cubic growth, when possible. Phenotypic, sire and dam correlations between body weights and hot standard carcass weight and rib eye muscle area were positive and moderate to high from birth to feedlot period. Management variation accounted for the largest proportion of total variation in both growth and carcass traits. Management correlations between carcass traits were high, except between rump fat depth and intramuscular fat (r = 0.26). Management correlations between body weight and carcass traits during the pre-weaning period were positive except for intramuscular fat. The correlations were low from birth to weaning, then increased dramatically and were high during the feedlot period.     

Surface display of organophosphorus hydrolase on E. coli using N-terminal domain of ice nucleation protein InaV

Recombinant Escherichia coli displaying organophosphorus hydrolase (OPH) was used to overcome the diffusion barrier limitation of organophosphorus pesticides. A new anchor system derived from the N-terminal domain of ice-nucleation protein from Pseudomonas syringae InaV (InaV-N) was used to display OPH onto the surface. The designed sequence was cloned in the vector pET-28a(+) and then was expressed in E. coli. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by InaV-N on the outer membrane, and the ability of recombinant E. coli to utilize diazinon as the sole source of energy, without growth inhibition, indicated its significant activity. The location of OPH was detected by comparing the activity of the outer membrane fraction with the inner membrane and cytoplasm fractions. Studies revealed that recombinant E. coli can degrade 50% of 2 mM chlorpyrifos in 2 min. It can be concluded that InaV-N can be used efficiently to display foreign functional protein, and these results highlight the high potential of an engineered bacterium to be used in bioremediation of pesticide-contaminated sources in the environment.     

Factors affecting the occurrence of postpartum prolonged luteal activity in clinically healthy high-producing dairy cows

The objective was to characterize risk factors affecting the occurrence of prolonged luteal phase (PLP) in postpartum, clinically healthy, high-producing dairy cows. Transrectal ultrasound examinations of the reproductive tract were performed twice weekly, from the 1st to 8th wk after calving in 151 multiparous clinically healthy lactating Holstein cows (mean ± SD of peak milk yield = 56.7 ± 7.4 kg). Serum samples were collected twice weekly to measure progesterone and every 2 wk to detect β-hydroxybutyrate (βHB), and α1-acid glycoprotein (AGP). Body condition score (BCS) was recorded weekly after calving. Based on the serum progesterone profile, 52 (34.4%) cows had normal ovarian activity (NLA), whereas 36 (23.8%) cows had a prolonged luteal phase (PLP), the most prevalent type of abnormal pattern of luteal activity. Furthermore, 63 cows with short luteal activity, delayed first ovulation, or cystic ovaries were excluded from this study. Serum AGP concentrations, as an indication of postpartum chronic endometritis, were not different (P > 0.05) between cows with NLA and PLP. Categories of peak milk yields (kg) were positively correlated with the incidence (%) of cows with PLP (r = 0.87, P = 0.02). Furthermore, milk yield peak, day of milk yield peak, mean milk yield (8 wk in milk), and milk yield on the expected day of luteolysis were higher (P < 0.05) in cows with PLP than NLA, and cows with PLP had greater loss of BCS (P = 0.007) than those with NLA. The likelihood of cows with PLP decreased by 0.9-fold for every 1 d delay of commencement of luteal activity (C-LA). Moreover, the likelihood of cows with PLP increased by 1.8-fold for each 1 mmol/L increase in the 1st wk serum βHB concentrations. In conclusion, higher mean of milk yield, greater BCS loss, earlier C-LA, and later peak milk yield were the major risk factors affecting the occurrence of postpartum PLP in clinically healthy, high-producing dairy cows.     

PloGO: plotting gene ontology annotation and abundance in multi-condition proteomics experiments

We describe the PloGO R package, a simple open-source tool for plotting gene ontology (GO) annotation and abundance information, which was developed to aid with the bioinformatics analysis of multi-condition label-free proteomics experiments using quantitation based on spectral counting. PloGO can incorporate abundance (raw spectral counts) or normalized spectral abundance factors (NSAF) data in addition to the GO annotation, as well as handle multiple files and allow for a targeted collection of GO categories of interest. Our main aims were to help identify interesting subsets of proteins for further analysis such as those arising from a protein data set partition based on the presence and absence or multiple pair-wise comparisons, as well as provide GO summaries that can be easily used in subsequent analyses. Though developed with label-free proteomics experiments in mind it is not specific to that approach and can be used for any multi-condition experiment for which GO information has been generated.     

Effects of whole body vibration on outer hair cells’ hearing response to distortion product otoacoustic emissions

Whole body vibration (WBV) is one of the most vexing problems in industries. There is a debate about the effect of WBV exposure on hearing system as vibration-induced hearing loss. The purpose of this study was to investigate outer hair cells' (OHCs') hearing response hearing response to distortion product otoacoustic emissions (DPOAEs) in rabbits exposed to WBV. It was hypothesized that the DPOAE response amplitudes (A(dp)) in rabbits exposed to WBV would be lower than those in control rabbits not exposed to WBV. New Zealand white (NZW) rabbits as vibration group (n = 6, exposed to WBV in the z-axis at 4-8 Hz and 1.0 ms(-2) root mean square for 8 h per day during five consecutive days) and NZW rabbits as control group (n = 6, not exposed to any WBV) were participated. A(dp) and noise floor levels (L(nf)) were examined on three occasions: day 0 (i.e., baseline), day 8 (i.e., immediately 1 h after exposure), and day 11 (i.e., 72 h following exposure) with f(2) frequencies ranging from 500 to 10,000 Hz and primaries L(1) and L(2) levels of 65 and 55 dB sound pressure level, respectively. Main effects were statistically found to be significant for group, time, and frequency (p < 0.05). DPOAE amplitudes were significantly larger for rabbits exposed to WBV, larger on day 8 and larger for mid to high f(2) frequencies (at and above 5,888.50 Hz). Main effects were not statistically found to be significant for ear (p > 0.05). Also, four statistically significant interactions including time by ear, time by frequency, group by frequency, and group by time were detected (p < 0.05). Contrary to the main hypothesis, DPOAE amplitudes were significantly larger for rabbits exposed to WBV. WBV exposure significantly led to enhanced mean A(dp) at mid to high frequencies rather than at low ones.