Isolation and characterization of 19 polymorphic microsatellite loci from the topmouth gudgeon, Pseudorasbora parva

The Asiatic topmouth gudgeon, Pseudorasbora parva, is recognized as one of the most invasive fish species in many countries outside of Asia. We isolated and characterized 19 microsatellite loci from P. parva. The polymorphism of these 19 loci was tested on 40 individuals of P. parva sampled from a wild population located in Ezhou, Hubei province of China. The loci had 5 to 11 alleles, with a mean of 7.7 at each locus; 11 loci conformed to Hardy-Weinberg equilibrium. The expected and observed heterozygosities ranged from 0.237 to 0.973 and from 0.647 to 0.914, respectively. All microsatellite loci were in linkage equilibrium. These microsatellite markers are potentially useful for the assessment of population genetic structure during invasion and dispersal of P. parva in new habitats.     

Validation of the ALS assay in adult patients with culture confirmed pulmonary tuberculosis

Background

We have earlier shown that Bacille Calmette-Guérin (BCG) vaccine-specific IgG Antibodies in Lymphocyte Supernatant (ALS) can be used for diagnosis of active tuberculosis (TB) in adults and children.

Methodology/Principal Findings

The ALS method was validated in a larger cohort (n = 212) of patients with suspicion of pulmonary TB using multiple antigens (BCG, LAM, TB15.3, TB51A, CFP10-ESAT6-A, CFP, CW) from Mycobacterium tuberculosis. The sensitivity and specificity of the ALS assay was calculated using non-TB patients as controls. The sensitivity and the specificity were highest with BCG vaccine (90% and 88% respectively) followed by LAM (89% and 87% respectively). Simultaneous assessment of multiple antigen-specific antibodies increased sensitivity (91%) and specificity (88%). Using higher lymphocyte count in smaller volume of culture media increased detection and reduced the assay duration to ∼30 hrs. Twenty one patients with clinical findings strongly suggestive of TB finally diagnosed as non-TB patients were positive by the ALS assay, of which 9 (43%) were positive for 7 antigens and 19 (90%) for at least 3 antigens.

Conclusions/Significance

Our findings show that simultaneous detection of antigens improves the diagnostic potential of the ALS assay; the modified method increases sensitivity and can provide results in <48 hours, and enable detection of some cases of pulmonary TB that are not detectable by standard methods.     

Secular citizenship and Muslim belonging in Turkey: migrant perspectives

In recent years, academic critique has worked toward exposing the underlying ethno-religious biases in official formulations of secular citizenship in Turkey. While such studies have highlighted the contradictions and inconsistencies that emerge in these grand narratives of national belonging, they have tended to privilege statist determinants of the bounds of nationhood. In this article, I offer an intervention into this discussion by exploring how citizens themselves encounter and embrace religion as a marker of national belonging in Turkey. In particular, I discuss the case of the 1950s Bosniak migration to Turkey and explore the salience of the migratory journey in informing their experiences of secular citizenship and national belonging.     

Structure and Community Composition of Sprout-Like Bacterial Aggregates in a Dinaric Karst Subterranean Stream

The Vjetrenica cave in the Dinaric Karst hosts a worldwide extraordinarily high cave biodiversity. Beside a diverse and specialized cave fauna, sprout-like formations attached to the bed of the cave stream were observed and described, but not further characterized, almost a century ago. Here we investigated these sprout-like microbial aggregates by the rRNA approach and detailed microscopy. Based on fluorescence in situ hybridization and ultrastructural analysis, the sprout-like formations are morphologically highly organized, and their core consists of a member of a novel deep-branching lineage in the bacterial phylum Nitrospirae. This organism displays an interesting cellular ultrastructure with different kinds of cytoplasmic inclusions and is embedded in a thick extracellular matrix, which contributes to the stability and shape of the aggregates. This novel bacterium has been provisionally classified as “Candidatus Troglogloea absoloni.” The surface of the sprout-like aggregates is more diverse than the core. It is colonized by a bacterial biofilm consisting primarily of filamentous Betaproteobacteria, whereas other microbial populations present in the crust include members of the Bacteriodetes, Gammaproteobacteria, Actinombacteria, Alphaproteobacteria, and Planctomycetes, which are intermingled with mineral inclusions. This study represents the first thorough molecular and ultrastructural characterization of the elusive sprout-like bacterial aggregates, which are also found in other cave systems of the Dinaric Karst. The discovery of Ca. Troglogloea absoloni contributes to the known biodiversity of subterranean ecosystems and especially of macroscopic structures formed in caves by microorganisms, whose composition and ecological function often remain enigmatic.     

Stripe rust analysis of D-genome synthetic wheats (2n = 6x = 42, AABBDD) and their molecular diversity

Stripe or yellow rust caused by Puccinia striiformis f. sp. tritici is a threat to many of the existing cultivars of Pakistan. Many attempts are being made to evolve new varieties resistant to stripe rust to reduce the losses caused by this disease. For this purpose, novel genes are needed to incorporate into the existing cultivars. These genes are found in the wild progenitors of wheat that are D-genome donors to wheat. As a result of extensive research, wheat synthetic hexaploids have been developed. These synthetics have resistances against biotic as well as abiotic stresses including the yellow rust. A group of such synthetics has been identified which seems resistant to this destructive disease. This group was tested under field conditions to identify resistance against stripe rust. The same population was analysed at molecular level to explore the genetic diversity for rust resistance. Genetic diversity among 34 selected synthetic hexaploid wheats was studied by random amplified polymorphic DNA (RAPD) analysis. A set of 12 RAPD primers was applied, and the level of polymorphism was found to be 46.67%. The coefficients in the range of 71–100% were detected by genetic similarity matrix based on Nei and Li's index. These coefficients were used for constructing a dendrogram using unweighted pair group of arithmetic means. Synthetic hexaploid line 34 was found to exhibit maximum genetic distances among the 34 selected lines. The same accession also showed excellent phenotypic characters with above average grain weight. These synthetic hexaploids carrying genetic potential for stripe rust resistance and morphological traits should be useful for improvement of existing wheat cultivars.     

Identification of interactions involved in the generation of nucleophilic reactivity and of catalytic competence in the catalytic site Cys/His ion pair of papain

Understanding the roles of noncovalent interactions within the enzyme molecule and between enzyme and substrate or inhibitor is an essential goal of the investigation of active center chemistry and catalytic mechanism. Studies on members of the papain family of cysteine proteinases, particularly papain (EC 3.4.22.2) itself, continue to contribute to this goal. The historic role of the catalytic site Cys/His ion pair now needs to be understood within the context of multiple dynamic phenomena. Movement of Trp177 may be necessary to expose His159 to solvent with consequent decrease in its degree of electrostatic solvation of (Cys25)-S(-). Here we report an investigation of this possibility using computer modeling of quasi-transition states and pH-dependent kinetics using 3,3'-dipyridazinyl disulfide, its n-propyl and phenyl derivatives, and 4,4'-dipyrimidyl disulfide as reactivity probes that differ in the location of potential hydrogen-bonding acceptor atoms. Those interactions that influence ion pair geometry and thereby catalytic competence, including by transmission of the modulatory effect of a remote ionization with pK(a) 4, were identified. A key result is the correlation between the kinetic influence of the modulatory trigger of pK(a) 4 and disruption of the hydrogen bond donated by the indole N-H of Trp177, the hydrophobic shield of the initial "intimate" ion pair. This hydrogen bond is accepted by the amide O of Gln19-a component of the oxyanion hole that binds the tetrahedral species formed from the substrate during the catalytic act. The disruption would be expected to contribute to the mobility of Trp177 and possibly to the effectiveness of the binding of the developing oxyanion.     

Sulf2 gene is alternatively spliced in mammalian developing and tumour tissues with functional implications

SULF2 enzyme regulates the activities of a number of signalling pathways that in many tissues are up-regulated during development and disease. As we recently showed for avian Sulf1, the present study demonstrates that mammalian Sulf2 gene can also generate functionally distinct splice variants that would regulate normal development and tumour growth differentially. It is thus important to distinguish SULF1/SULF2 isoforms in mammalian tissues to understand their functional and clinical relevance to disease. This study demonstrates that unlike normal adult lung with little or no SULF2 expression, this enzyme is expressed at high levels in most lung tumours showing differential cellular distribution of full length and shorter SULF2 variants in such tumours. Furthermore, we show that the short SULF2 splice variants are associated with those signalling pathways that are inhibited by full length SULF1/SULF2 variants and therefore could promote growth in such lung tumours.     

The β-blocker Nebivolol Is a GRK/β-arrestin Biased Agonist

Nebivolol, a third generation β-adrenoceptor (β-AR) antagonist (β-blocker), causes vasodilation by inducing nitric oxide (NO) production. The mechanism via which nebivolol induces NO production remains unknown, resulting in the genesis of much of the controversy regarding the pharmacological action of nebivolol. Carvedilol is another β-blocker that induces NO production. A prominent pharmacological mechanism of carvedilol is biased agonism that is independent of Gα_s and involves G protein-coupled receptor kinase (GRK)/β-arrestin signaling with downstream activation of the epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK). Due to the pharmacological similarities between nebivolol and carvedilol, we hypothesized that nebivolol is also a GRK/β-arrestin biased agonist. We tested this hypothesis utilizing mouse embryonic fibroblasts (MEFs) that solely express β₂-ARs, and HL-1 cardiac myocytes that express β₁- and β₂-ARs and no detectable β₃-ARs. We confirmed previous reports that nebivolol does not significantly alter cAMP levels and thus is not a classical agonist. Moreover, in both cell types, nebivolol induced rapid internalization of β-ARs indicating that nebivolol is also not a classical β-blocker. Furthermore, nebivolol treatment resulted in a time-dependent phosphorylation of ERK that was indistinguishable from carvedilol and similar in duration, but not amplitude, to isoproterenol. Nebivolol-mediated phosphorylation of ERK was sensitive to propranolol (non-selective β-AR-blocker), AG1478 (EGFR inhibitor), indicating that the signaling emanates from β-ARs and involves the EGFR. Furthermore, in MEFs, nebivolol-mediated phosphorylation of ERK was sensitive to pharmacological inhibition of GRK2 as well as siRNA knockdown of β-arrestin 1/2. Additionally, nebivolol induced redistribution of β-arrestin 2 from a diffuse staining pattern into more intense punctate spots. We conclude that nebivolol is a β₂-AR, and likely β₁-AR, GRK/β-arrestin biased agonist, which suggests that some of the unique clinically beneficial effects of nebivolol may be due to biased agonism at β₁- and/or β₂-ARs.     

Multi-strain Inoculation with PGPR Producing ACC Deaminase is More Effective Than Single-strain Inoculation to Improve Wheat (<i>Triticum aestivum</i>) Growth and Yield

Rhizosphere bacteria that colonize plant roots and confer beneficial effects are referred as plant growth promoting rhizobacteria (PGPR). Among all PGPR, some rhizobacteria have an ability to produce ACC deaminase enzyme. This enzyme catalyzes stress ACC into a-ketobutyrate and ammonia instead of letting it to be converted to ethylene. Ethylene level rises in plants under stress conditions i.e., drought, salinity, poor soil fertility etc. As poor soil fertility is a big hurdle to achieve the optimum yield of crops, inoculation of ACC deaminase PGPR can overcome this problem to some extent. The aim of the current study was to examine the influence of multi-strain and single-strain inoculation of different ACC deaminase producing PGPR on wheat growth and yield. There were three PGPR strains, Enterobacter cloacae, Serratia ficaria and Burkholderia phytofirmans which were used as consortia and single-strain inoculations. The results showed that inoculation of E. cloacae + S. ficaria + B. phytofirmans significantly increased plant height (63%), spike length (61%), number of spikelets spike^-1 (61%), number of grains spike-1 (131%), 1000 grains weight (33%), grains yield (71%), straw yield (71%) and biological yield (68%) of wheat as compared to control. A significant improvement in N (37 and 200%), P (46 and 166%) and K (39 and 61%) of seeds and shoot respectively, validated the efficacious and more effective role of multi-strain (E. cloacae + S. ficaria + B. phytofirmans) inoculation over control. It is obviously concluded that multi-strain ACC deaminase producing PGPR inoculation is a better approach as compared to singlestrain inoculation for the improvement in growth and yield of wheat.