Serotype Diversity of Astroviruses in Rawalpindi,Pakistan during 2009–2010

Astroviruses are globally known enteropathogens causing gastroenteritis and diarrhea, with eight well defined serotypes. Epidemiological studies have recognized serotype-1 as the most common subtype but no such data is available in Pakistan. During 2009–2010, we found astroviruses in 41 out of 535 (7%) samples collected from hospitalized children. Thirty one strains belonged to serotype-1 and clustered into two distinct lineages. Serotype-3, -4 and -6 were detected with 97–98% genetic homology to Indian and Chinese strains.     

Characterization of a Novel Enterovirus Serotype and an Enterovirus EV-B93 Isolated from Acute Flaccid Paralysis Patients

Non-polio enteroviruses (NPEVs) are among the most common viruses infecting humans worldwide. Most of these infections are asymptomatic but few can lead to systemic and neurological disorders like Acute Flaccid Paralysis (AFP). Acute Flaccid Paralysis is a clinical syndrome and NPEVs have been isolated frequently from the patients suffering from AFP but little is known about their causal relationship. The objective of this study was to identify and characterize the NPEV serotypes recovered from 184 stool samples collected from AFP patients in Federally Administered Tribal Areas (FATA) in north-west of Pakistan. Overall, 44 (95.6 %) isolates were successfully typed through microneutralization assay as a member of enterovirus B species including echovirus (E)-2, E-3, E-4, E-6, E-7, E-11, E-13, E-14, E-21 and E-29 while two isolates (PAK NIH SP6545B and PAK NIH SP1202B) remained untypeable. The VP1 and capsid regions analysis characterized these viruses as EV-B93 and EV-B106. Phylogenetic analysis confirmed that PAK NIH isolates had high genetic diversity and represent distinct genotypes circulating in the country. Our findings highlight the role of NPEVs in AFP cases to be thoroughly investigated especially in high disease risk areas, with limited surveillance activities and health resources.     

Development and utilization of an InDel marker linked to the fertility restorer genes of CMS-D8 and CMS-D2 in cotton

Molecular Biology Reports - The cytoplasmic male sterility (CMS) system is a useful tool for commercial hybrid cotton seed production. Two main CMS systems, CMS-D8 and CMS-D2, have been recognized...     

Genetic diversity of Paramecium species on the basis of multiple loci analysis and ITS secondary structure models

Among ciliates, Paramecium has become a privileged model for the study of “species problem” particularly in the case of the “Paramecium aurelia complex” that has been intensely investigated. Despite extensive studies, the taxonomy of Paramecium is still challenging. The major problem is an uneven sampling of Paramecium with relatively few representatives of each species. To investigate species from the less discovered region (Pakistan), 10 isolates of Paramecium species including a standing-alone FT8 strain previously isolated by some of us were subjected to molecular characterization. Fragments of 18S recombinant DNA (rDNA), ITS1-5.8S-ITS2-5′LSU rDNA, cytochrome c oxidase subunit II, and hsp70 genes were used as molecular markers for phylogenetic analysis of particular isolates. The nucleotide sequences of polymerase chain reaction products of all markers were compared with the available sequences of relevant markers of other Paramecium species from GenBank. Phylogenetic trees based on all molecular markers showed that all the nine strains had a very close relationship with Paramecium primaurelia except for the FT8 strain. FT8 consistently showed its unique position in comparison to all other species in the phylogenetic trees. Available sequences of internal transcribed spacer 1 (ITS1) and ITS2 and some other ciliate sequences from GenBank were used for the construction of secondary models. Two highly conserved helices supported by compensatory base changes among all ciliates of ITS2 secondary structures were found similar to other eukaryotes. Therefore, the most conserved 120 to 180 base pairs regions were identified for their comparative studies. We found that out of the three helices in ITS1 structure, helix B was more conserved in Paramecium species. Despite various substitutions in the primary sequence, it was observed that secondary structures of ITS1 and ITS2 could be helpful in interpreting the phylogenetic relationships both at species as well as at generic level.     

A micropropagation protocol for <Emphasis Type="Italic">Cassia angustifolia</Emphasis> Vahl. from root explants

The aim of this study was to develop a new micropropagation system for Cassia angustifolia Vahl., an important medicinal legume using root explant as starting material. Root explants taken from 30-day-old aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators: 6-benzyladenine (BA), kinetin (Kn), and thidiazuron (TDZ). Organogenic nodular calli obtained on MS + TDZ (1.0 μM) were transferred to shoot regeneration medium supplemented with different cytokinins (BA, Kn or TDZ) either alone or in combination with auxin:indole-3-acetic acid or α-naphthalene acetic acid. Maximum shoot regeneration frequency (90%) was obtained on MS + BA (2.5 μM) + NAA (0.6 μM) wherein a maximum of 42.76 ± 1.47 shoot buds per explant were induced with a maximum conversion rate of 35.63 ± 0.75 shoots per explant and average shoot length of 5.43 ± 0.20 cm. Elongated microshoots were successfully rooted under ex vitro conditions by pulse treatment in 200 μM of indole-3-butyric acid for half an hour. Microshoots were rooted, acclimatized and hardened off simultaneously in sterilized soilrite inside the growth room and then established in pots containing sterilized soil and manure (1:1) and grown under greenhouse condition with 90% survival rate. The histological sections at different developmental stages of shoot buds revealed the organization of nodular meristematic zone leading to the orientation and differentiation of shoot buds in large number and thereafter conversion into healthy shoots.     

E6 and E7 from human papillomavirus type 16 cooperate to target the PDZ protein Na/H exchange regulatory factor 1

Previous studies have shown that the PDZ-binding motif of the E6 oncoprotein from the mucosal high-risk (HR) human papillomavirus (HPV) types plays a key role in HPV-mediated cellular transformation in in vitro and in vivo experimental models. HR HPV E6 oncoproteins have the ability to efficiently degrade members of the PDZ motif-containing membrane-associated guanylate kinase (MAGUK) family; however, it is possible that other PDZ proteins are also targeted by E6. Here, we describe a novel interaction of HPV type 16 (HPV16) E6 with a PDZ protein, Na(+)/H(+) exchange regulatory factor 1 (NHERF-1), which is involved in a number of cellular processes, including signaling and transformation. HPV16 E6 associates with and promotes the degradation of NHERF-1, and this property is dependent on the C-terminal PDZ-binding motif of E6. Interestingly, HPV16 E7, via the activation of the cyclin-dependent kinase complexes, promoted the accumulation of a phosphorylated form of NHERF-1, which is preferentially targeted by E6. Thus, both oncoproteins appear to cooperate in targeting NHERF-1. Notably, HPV18 E6 is not able to induce NHERF-1 degradation, indicating that this property is not shared with E6 from all HR HPV types. Downregulation of NHERF-1 protein levels was also observed in HPV16-positive cervical cancer-derived cell lines, such as SiHa and CaSki, as well as HPV16-positive cervical intraepithelial neoplasia (CIN). Finally, our data show that HPV16-mediated NHERF-1 degradation correlates with the activation of the phosphatidylinositol-3'-OH kinase (PI3K)/AKT signaling pathway, which is known to play a key role in carcinogenesis.     

Effects of Plant Growth Regulators on Seed Filling,Endogenous Hormone Contents and Maize Production in Semiarid Regions

In this study, we determined whether the application of uniconazole alone or combined with ethephon could enhance the seed-filling rates in maize by regulating the endogenous hormone contents. Uniconazole was applied to the foliage at concentrations of 0 (CK), 25 (U₂₅), 50 (U₅₀) and 75 (U₇₅) mg L⁻¹ at the 12-leaf stage. In addition, uniconazole was applied to the foliage at the 12-leaf stage and ethephon at 10 days after silking stage at concentrations of 0 (CK), 25 mg L⁻¹ uniconazole + 100 mg L⁻¹ ethephon (U₂₅ + E₁₀₀), 50 mg L⁻¹ uniconazole + 200 mg L⁻¹ ethephon (U₅₀ + E₂₀₀) and 75 mg L⁻¹ uniconazole + 300 mg L⁻¹ ethephon (U₇₅ + E₃₀₀). Uniconazole applied alone or in combination with ethephon significantly improved ear characters and grain yield. Uniconazole applied alone or combination with ethephon significantly improved the dry matter accumulation in seeds and seed-filling rates. Uniconazole significantly increased the abscisic acid (ABA) and zeatin (Z) + zeatin riboside (ZR) contents of seeds, but reduced the gibberellic acid (GA) contents. The application of uniconazole combined with ethephon decreased the ABA, Z + ZR and GA contents in seeds. The ABA and Z + ZR contents were significantly positively correlated, whereas the GA content was negatively correlated with the maximum seed weight, maximum seed-filling rate and mean seed-filling rate. The application of uniconazole alone significantly improved the seed-filling rates in maize by regulating the endogenous hormone contents. Thus, we conclude that uniconazole application at 50 mg L⁻¹ in the 12-leaf stage can enhance the maize production.

     

Seed priming improves the emergence potential,growth and antioxidant system of Moringa oleifera under saline conditions

Moringa oleifera is a multipurpose plant which is now being promoted as a fodder crop. The present study was conducted to induce the tolerance in moringa plants to emerge and grow under saline conditions. For this, moringa seeds were primed with aerated water (hydropriming) and moringa leaf extract (MLE) for 12 and 24 h and studied for its emergence, potential growth behaviour, mineral composition, chlorophyll contents and antioxidant activities in comparison with unprimed seeds to investigate the physiological changes in moringa plants under saline conditions. The seeds were sown in plastic pots filled with acid washed sand at four salinity levels (3, 6, 10, 14 dS m^?1) in a completely randomized design with three replications. It was found that salinity >6 dS m^?1 reduced the emergence, growth and vigour of moringa plants but hydropriming (12 h) enhanced moringa emergence at 10 dS m^?1 followed by MLE priming (12 h). Maximum aboveground biomass and photosynthetic pigments were recorded when the seeds were hydroprimed (12 h) but maximum root length and number of roots were found in MLE primed (12 h) moringa plants. Significant decrease in K⁺:Na⁺ ratio with increasing salinity levels resulted in low K⁺ and Mg²⁺ uptake and Na⁺ toxicity in moringa leaves which resulted in reduced chlorophyll contents at 14 dS m^?1 but a significant increase in chlorophyll a and b contents and total phenolics were found in hydroprimed seeds (12 h) while the antioxidant activities of superoxide dismutase, peroxidase and catalas were improved by MLE priming (12 h). This study concludes that moringa emergence and growth performance can be improved by hydropriming under saline conditions.     

Markers/parameters for the evaluation of natural resistance status of small ruminants against gastrointestinal nematodes

The high prevalence of anthelmintic-resistant gastrointestinal nematodes (GINs) throughout the world has led to the need for alternative worm control strategies. One of the possible substitutes to reduce the problems of drug resistance and residue is the evaluation/breeding of small ruminants for greater resistance to the GINs (organically produced), which in turn would be a helpful tool to predict the performance of an animal. At present, the existing diversity in the genetic potential to resist/tolerate GINs infection both within and between breeds has been validated. Successful selection of animals to define the genotype and identified resistance is related to the employed markers. A number of phenotypic traits such as faecal egg count (FEC), worm burden, serum antibodies, peripheral eosinophilia, packed cell volume, live weight, serum protein and albumin concentrations have been used for this purpose both in natural and artificial infections. Relatively resistant/tolerant animals have also been found to have mastocytosis, globule leucocytes, high levels of histamine and immunoglobulin (Ig) A and IgE concentrations. Of these traits, the principal and most practical measurement used to assess resistance status in animals undergoing similar parasite challenges is FEC. FEC has a positive/negative correlation with other biochemical, cellular and immunological parameters; however, the reliability of individual trial is often questioned and valuable information regarding the genetic makeup can be obtained from pooled data of a large number of trials and parameters. This paper covers all the aspects reported in the literature on various parameters considered to evaluate the resistance status of a range of small ruminant breeds.