Development of male sterility by silencing Bcp1 gene of Arabidopsis through RNA interference

The development of male sterility is one of the most important steps in hybrid seed production. Several methods for the abortion of pollens based on conventional as well as genetic engineering are reported for the various crop species. Here we have investigated the use of RNA interference (RNAi) technology to silence a male specific gene, Bcp1 in the model host Arabidopsis thaliana. Bcp1 is active in both diploid tapetum and haploid microspores. Three batches of explants (A. thaliana) were selected onherbicide glufosinate ammonium and putative transgenes were confirmed through PCR and Southern hybridization. About 49 out of 58 Arabidopsis lines transformed with RNAi construct containing Bcp1 sequences were male sterile. Transgenic plants were phenotypically indistinguishable from nontransgenic plants and by crossing with non-transgenic fertile pollens successful seed set was observed. The Bcp1 gene was also amplified from chilies, tomato and brassica. The present study resulted in developing male sterile A. thaliana (Eco. Columbia) line through genetic engineering. The diversity of Bcp1 gene will help us to produce transgenic male sterile plants in crop plants

Identification of an N-terminal 27 kDa fragment of Mycoplasma pneumoniae P116 protein as specific immunogen in M. pneumoniae infections

<p>Abstract</p> <p>Background</p> <p><it>Mycoplasma pneumoniae </it>is an important cause of respiratory tract infection and is increasingly being associated with other diseases such as asthma and extra-pulmonary complications. Considerable cross-reactivity is known to exist between the whole cell antigens used in the commercial serological testing assays. Identification of specific antigens is important to eliminate the risk of cross-reactions among different related organisms. Adherence of <it>M. pneumoniae </it>to human epithelial cells is mediated through a well defined apical organelle to which a number of proteins such as P1, P30, P116 and HMW1-3 have been localized, and are being investigated for adhesion, gliding and immunodiagnostic purposes.</p> <p>Methods</p> <p>A 609 bp fragment P116_(N-27),corresponding to the N-terminal region of <it>M. pneumoniae </it>P116 gene was cloned and expressed. A C-terminal fragment P1_(C-40),of P1 protein of <it>M. pneumoniae </it>was also expressed. Three IgM ELISA assays based on P116_(N-27),P1_(C-40)and (P116 _(N-27)+ P1_(C-40)) proteins were optimized and a detailed analysis comparing the reactivity of these proteins with a commercial kit was carried out. Comparative statistical analysis of these assays was performed with the SPSS version 15.0.</p> <p>Results</p> <p>The expressed P116_(N-27)protein was well recognized by the patient sera and was immunogenic in rabbit. P1_(C-40)of <it>M. pneumoniae </it>was also immunogenic in rabbit. In comparison to the reference kit, which is reported to be 100% sensitive and 75% specific, ELISA assay based on purified P116_(N-27),P1_(C-40)and (P116_(N-27)+ P1_(C-40)) proteins showed 90.3%, 87.1% and 96.8% sensitivity and 87.0%, 87.1% and 90.3% specificity respectively. The p value for all the three assays was found to be < 0.001, and there was a good correlation and association between them.</p> <p>Conclusion</p> <p>This study shows that an N-terminal fragment of P116 protein holds a promise for serodiagnosis of <it>M. pneumoniae </it>infection. The IgM ELISA assays based on the recombinant proteins seem to be suitable for the use in serodiagnosis of acute <it>M. pneumoniae </it>infections. The use of short recombinant fragments of P116 and P1 proteins as specific antigens may eliminate the risk of cross-reactions and help to develop a specific and sensitive immunodiagnostic assay for <it>M. pneumoniae </it>detection.</p

Delay in diagnosis of tuberculosis in Rawalpindi, Pakistan

<p>Abstract</p> <p>Background</p> <p>Delay in diagnosis and treatment of tuberculosis (TB) may enhance the chances of morbidity and mortality and play a key role in continuous transmission of the bacilli. The objective of this study was to describe health care seeking behavior of suspected TB patients and initial diagnostic work up prior to consultation and diagnosis at National TB Center (NTC).</p> <p>Findings</p> <p>Interviews of 252 sputum smear positive patients were taken from NTC, Rawalpindi. The duration between on-set of symptoms and start of treatment was considered as the total delay and correlated with general characteristics of TB patients. The proportion of males and females were 49.6% and 50.4% with median age of 25 and 24 years respectively. A median delay of 56 days (8 weeks) was observed which was significantly associated with age, cough and fever. More than 50% of the current patients had a history of contact with previously diagnosed TB patients. The majority of patients (63%) visited health care providers within three weeks of appearance of symptoms but only thirty five percent were investigated for TB diagnosis.</p> <p>Conclusion</p> <p>Cough and fever are being ignored as likely symptoms of TB by patients as well as health care providers resulting in delay. Engaging private practitioners through public private mix (PPM) approach for expansion of TB diagnosis and increasing public awareness could be more beneficial to reduce delay.</p

Impact of different levels of zinc and nitrogen on growth, productivity, and quality of aromatic rice cultivated under various irrigation regimes in two districts of Pakistan

Rice is a staple food for more than 50% of the global population and it is one of the most valuable cereal crops. To fulfill the dietary requirement of the ever-growing world population, an increase in per-unit production of rice is direly required. In Pakistan, it stands as the 2nd in consumption after wheat, which is a staple food. A huge gap is observed between yield potential and actual yield of the aromatic rice cultivars at a farmer-field level. The significant limitations responsible for this gap are shortage of irrigation water, inappropriate application of fertilizers, less plant population, deficiency of micronutrients, and improper and poor plant protection measures. A field study was planned to assess the yield response and quality attributes of aromatic rice to three levels of zinc (Zn) and nitrogen (N) under three irrigation regimes (8-, 12-, and 16-acre inches) in the Sheikhupura and Sargodha districts of Pakistan. Irrigation treatments significantly influenced the growth, yield, and quality attributes; however, maximum improvement was observed by the application of irrigation at 12-acre inches. Among the Zn treatments, application of Zn at 10 kg ha–1 was observed to be more responsive to improving the growth and quality parameters of aromatic rice crops. In the case of N treatments, application of N at 140 kg ha–1 produced the maximum total tillers, as well as productive tillers per hill, spikelets per panicle, leaf area index, leaf area duration, crop growth rate, total dry matter, harvest index, kernel length, kernel width, and 1,000-kernel weight. Application of N at 140 kg ha–1 not only improved the growth attributes but also increased the net assimilation rate, photosynthetically active radiation, and radiation use efficiency, with respect to total dry matter and kernel yield. The maximum percentage of normal kernels and minimum percentage of opaque, abortive, and chalky kernels were also recorded by application of N at 140 kg ha–1. The outcomes of current experiments depicted that application of irrigational water, zinc, and nitrogen at 12-acre inches, 10, and 140 kg ha–1, respectively, are responsible to achieve maximum resource utilization efficiency, along with increased yield and quality of rice

Molecular markers-based DNA fingerprinting coupled with morphological diversity analysis for prediction of heterotic grouping in sunflower (Helianthus annuus L.)

Cultivated sunflower holds a very narrow genetic base and the efficient utilization of available genetic diversity is very important for a successful breeding program. In the present study, 109 sunflower genotypes were assessed for diversity paneling through a combined approach of morphological and molecular markers analysis. Morphological parameters including days to flower initiation, days to flower completion, plant height, stem curvature, number of leaves per plant, leaf area, head diameter, hundred seed weight, and seed yield per plant were studied. Simple sequence repeats (40 DNA markers) were deployed for diversity profiling. Data were analyzed by both univariate and multivariate statistics. SD and coefficient of variation confirm the presence of significant amounts of genetic variation for all the morphological parameters. Cluster Analysis and Principal Component Analysis further confirm the presence of distinct grouping patterns in the studied material. Cluster analysis of both morphological and molecular analysis revealed that restorer lines tend to group separately from A, B, and open-pollinated lines. Further grouping, at the sub-cluster level, revealed six distinct sub-clusters in each of the two major clusters. In total, 12 genotypes, 6 CMS lines (CMS-HAP-12, CMS-HAP-54, CMS-HAP-56, CMS-HAP-99, CMS-HAP-111, and CMS-HAP-112) and 6 restorer lines (RHP-38, RHP-41, RHP-53, RHP-68, RHP-69, and RHP-71) could be used as potential parents for hybrid development. As genotypes of similar genetic backgrounds tend to group closer, it is deduced that one genotype with the highest seed yield per plant could be used for further hybrid breeding programs in sunflowers

Secure Virtual Mobile Small Cells: A Stepping Stone Towards 6G

YesAs 5th Generation research reaches the twilight, the research community must go beyond 5G and look towards the 2030 connectivity landscape, namely 6G. In this context, this work takes a step towards the 6G vision by proposing a next generation communication platform, which aims to extend the rigid coverage area of fixed deployment networks by considering virtual mobile small cells (MSC) that are created on demand. Relying on emerging computing paradigms such as NFV (Network Function Virtualization) and SDN (Software Defined Networking), these cells can harness radio and networking capability locally reducing protocol signalling latency and overhead. These MSCs constitute an intelligent pool of networking resources that can collaborate to form a wireless network of MSCs providing a communication platform for localized, ubiquitous and reliable connectivity. The technology enablers for implementing the MSC concept are also addressed in terms of virtualization, lightweight wireless security, and energy efficient RF. The benefits of the MSC architecture towards reliable and efficient cell-offloading are demonstrated as a use-case.This project has received funding from the European Union's H2020 research and innovation program under grant agreement H2020-MCSAITN- 2016-SECRET 722424 [2]

Cholangiocyte organoids can repair bile ducts after transplantation in the human liver.

Organoid technology holds great promise for regenerative medicine but has not yet been applied to humans. We address this challenge using cholangiocyte organoids in the context of cholangiopathies, which represent a key reason for liver transplantation. Using single-cell RNA sequencing, we show that primary human cholangiocytes display transcriptional diversity that is lost in organoid culture. However, cholangiocyte organoids remain plastic and resume their in vivo signatures when transplanted back in the biliary tree. We then utilize a model of cell engraftment in human livers undergoing ex vivo normothermic perfusion to demonstrate that this property allows extrahepatic organoids to repair human intrahepatic ducts after transplantation. Our results provide proof of principle that cholangiocyte organoids can be used to repair human biliary epithelium

Disorders of sex development : insights from targeted gene sequencing of a large international patient cohort

Background: Disorders of sex development (DSD) are congenital conditions in which chromosomal, gonadal, or phenotypic sex is atypical. Clinical management of DSD is often difficult and currently only 13% of patients receive an accurate clinical genetic diagnosis. To address this we have developed a massively parallel sequencing targeted DSD gene panel which allows us to sequence all 64 known diagnostic DSD genes and candidate genes simultaneously. Results: We analyzed DNA from the largest reported international cohort of patients with DSD (278 patients with 46, XY DSD and 48 with 46, XX DSD). Our targeted gene panel compares favorably with other sequencing platforms. We found a total of 28 diagnostic genes that are implicated in DSD, highlighting the genetic spectrum of this disorder. Sequencing revealed 93 previously unreported DSD gene variants. Overall, we identified a likely genetic diagnosis in 43% of patients with 46, XY DSD. In patients with 46, XY disorders of androgen synthesis and action the genetic diagnosis rate reached 60%. Surprisingly, little difference in diagnostic rate was observed between singletons and trios. In many cases our findings are informative as to the likely cause of the DSD, which will facilitate clinical management. Conclusions: Our massively parallel sequencing targeted DSD gene panel represents an economical means of improving the genetic diagnostic capability for patients affected by DSD. Implementation of this panel in a large cohort of patients has expanded our understanding of the underlying genetic etiology of DSD. The inclusion of research candidate genes also provides an invaluable resource for future identification of novel genes

Reclaiming democracy in the square? Interpreting the movements of 2011-12

In this report we examine the demands and aspirations, for greater democracy, rule of law, social justice, and dignity and against inequality and corruption that brought people into the streets and squares across the globe in 2011-12