BIOLOGICAL CHANGES OCCURED IN SOYBEAN SEED DURING EXPOSING TO SEVERAL TYPES OF SEED PRIMING

Biological experiments were carried out at Agronomy Seed Lab., Agronomy Dept., Faculty of Agriculture, Ain Shams University and Ain Shams Center For Genetic Engineering and Biotechnology ACGEB, Genetic Dept., Faculty of Agriculture, Ain Shams University during 2016/2017 season. Priming and its duration were investigated whereas different types of priming (hydro-priming, osmopriming and salt-priming) and different periods of each type (short- medium and long periods) were stuied. Newly harvested soybean seeds cultivar (Giza 111) were submitted from Field Crop Institute, Agricultural Research Center (ARC). It was found that type of priming enhanced germination percentage significantly from low performance of 40% to 51%, 68% and 75.5% for hydration, osmopriming and salt priming respectively. Extending exposing period to the longest period gave significantly maximum increment in seed germination. Maximum germination enhancement was achieved when calcium chloride solution was used for the longest period (48 hrs.) giving value of 96.0%. Such increment reached 140% as compared with control.  It was noticeable that increasing soaking period to the longest period examined in this investigation accelerated the rate of germination to a maximum level. Salt priming produced longest soybean seedling shoot when compared with control. Overall, for most results obtained in this trial,   seed primed with CaCl2 showed better perfor mance than those primed with water or PEG solution.  Seedling dry weight revealed a significant effect in a similar manner of seedling length. The longest exposing priming period showed a significant effect on seedling dry weight. Soybeans seeds proteins exposed to priming for all three periods used in this investigation varied from control, whereas number of protein bands on SDS gel increased from 10 bands separated on control pattern to 13, 11 and 12 for hydration treatment at periods of 6, 12 and 24 hrs., respectively. Also number of mono poly-uni. and unique bands varied as well as its intensive dye which reflect that amount of protein formed varied between treatments.  Considering exposing seeds to salt solution of CaCl2 for longest period (48 hrs.) less bands of separated protein were formed on SDS gel. Biological seed quality was assessed by extracting proteins on native polyacrylamide gel electrophoresis, whereas, all priming treatment at application periods caused on increasing in peroxidase activity compared to control (untreated seeds).  It was remarkably that the longest period of expose showed the highest peroxidase activity when compare to control and also for the short and medium exposing periods (6 and 12 hrs.). Also it was noticed that there were a unique diffuse band at the end of the lane, these bands were less intensive in staining color, meaning that their activities is less than those extracted from hydro-primed seeds for short and medium periods (6 and 12 hrs. respectively).  Seed esterase activity bands had two prolonged regions, these bands appeared as a diffuse bands. Esterase activity bands of PAGE gel showed less activity as subjected to all priming types at any period used in this investigation (long, medium and short) when compared to untreated seed (control). It was clear that increasing period of exposing seeds to priming reduced esterase activity since the intensive band diminished in their intensive color

Genetic Engineering of Schizosaccharomyces pombe to Produce Bacterial Polyhydroxyalkanotes

A commercial use of microbial produced products, like polyhydroxyalkanotes (PHAs), in the sense of an environmental precaution appears meaningful and necessary. In order to more economically produce microbial products, this investigation was focused on suitable producers, like the yeast Schizosaccharomyces pombe. Since it is not capable of the PHA synthesis, easily cultured and they must be modified genetically. Therefore, the genes of the phb biosynthesis pathway of Ralstonia eutropha [beta-ketothiolase (phbARe); acetoacetyl-CoA reductase (phbBRe); as well as phb synthase (phbCRe), located onto the plasmid pBHR68 were cloned into the cohesive ended pYIplac128 integrated vector that transformed into the chromosome of the yeast Schizosaccharomyces pombe strain Q01. Under the optimized cultivation conditions, the transgenic yeast S. pombe strain Q01/phb was able to produce phb and accumulated up to 9.018 % phb. The presence of heterologous DNA in the transgenic yeast was examined by means of Western blot analysis. In addition, both PHA synthase activity and kinetics were determined. The UV/Vis, 1H and 13C NMR spectral analysis have confirmed that the polymer produced by the yeast S. pombe strain Q01/phb is a pure homopolymer of 3-hydroxybutyric acid

Genomic characterization of a newly discovered coronavirus associated with acute respiratory distress syndrome in humans

A novel human coronavirus (HCoV-EMC/2012) was isolated from a man with acute pneumonia and renal failure in June 2012. This report describes the complete genome sequence, genome organization, and expression strategy of HCoV-EMC/2012 and its relation with known coronaviruses. The genome contains 30,119 nucleotides and contains at least 10 predicted open reading frames, 9 of which are predicted to be expressed from a nested set of seven subgenomic mRNAs. Phylogenetic analysis of the replicase gene of coronaviruses with completely sequenced genomes showed that HCoV-EMC/2012 is most closely related to Tylonycteris bat coronavirus HKU4 (BtCoV-HKU4) and Pipistrellus bat coronavirus HKU5 (BtCoV-HKU5), which prototype two species in lineage C of the genus Betacoronavirus. In accordance with the guidelines of the International Committee on Taxonomy of Viruses, and in view of the 75% and 77% amino acid sequence identity in 7 conserved replicase domains with BtCoVHKU4 and BtCoV-HKU5, respectively, we propose that HCoV-EMC/2012 prototypes a novel species in the genus Betacoronavirus. HCoV-EMC/2012 may be most closely related to a coronavirus detected in Pipistrellus pipistrellus in The Netherlands, but

Human coronavirus EMC does not require the SARS-coronavirus receptor and maintains broad replicative capability in mammalian cell lines

A new human coronavirus (hCoV-EMC) has emerged very recently in the Middle East. The clinical presentation resembled that of the severe acute respiratory syndrome (SARS) as encountered during the epidemic in 2002/2003. In both cases, acute renal failure was observed in humans. HCoV-EMC is a member of the same virus genus as SARS-CoV but constitutes a sister species. Here we investigated whether it might utilize angiotensin-converting enzyme 2 (ACE2), the SARS-CoV receptor. Knowledge of the receptor is highly critical because the restriction of the SARS receptor to deep compartments of the human respiratory tract limited the spread of SARS. In baby hamster kidney (BHK) cells, lentiviral transduction of human ACE2 (hACE2) conferred permissiveness and replication for SARS-CoV but not for hCoV-EMC. Monkey and human kidney cells (LLC-MK2, Vero, and 769-P) and swine kidney cells were permissive for both viruses, but only SARS-CoV infection could be blocked by anti-hACE2 antibody and could be neutralized by preincubation of virus with soluble ACE2. Our data show that ACE2 is neither necessary nor sufficient for hCoV-EMC replication. Moreover, hCoV-EMC, but not SARS-CoV, replicated in cell lines from Rousettus, Rhinolophus, Pipistrellus, Myotis, and Carollia bats, representing four major chiropteran families from both suborders. As human CoV normally cannot replicate in bat cells from different families, this suggests that hCoV-EMC might use a receptor molecule that is conserved in bats, pigs, and humans, implicating a low barrier against cross-host transmission

Mapping child growth failure across low- and middle-income countries

Childhood malnutrition is associated with high morbidity and mortality globally1. Undernourished children are more likely to experience cognitive, physical, and metabolic developmental impairments that can lead to later cardiovascular disease, reduced intellectual ability and school attainment, and reduced economic productivity in adulthood2. Child growth failure (CGF), expressed as stunting, wasting, and underweight in children under five years of age (0�59 months), is a specific subset of undernutrition characterized by insufficient height or weight against age-specific growth reference standards3�5. The prevalence of stunting, wasting, or underweight in children under five is the proportion of children with a height-for-age, weight-for-height, or weight-for-age z-score, respectively, that is more than two standard deviations below the World Health Organization�s median growth reference standards for a healthy population6. Subnational estimates of CGF report substantial heterogeneity within countries, but are available primarily at the first administrative level (for example, states or provinces)7; the uneven geographical distribution of CGF has motivated further calls for assessments that can match the local scale of many public health programmes8. Building from our previous work mapping CGF in Africa9, here we provide the first, to our knowledge, mapped high-spatial-resolution estimates of CGF indicators from 2000 to 2017 across 105 low- and middle-income countries (LMICs), where 99 of affected children live1, aggregated to policy-relevant first and second (for example, districts or counties) administrative-level units and national levels. Despite remarkable declines over the study period, many LMICs remain far from the ambitious World Health Organization Global Nutrition Targets to reduce stunting by 40 and wasting to less than 5 by 2025. Large disparities in prevalence and progress exist across and within countries; our maps identify high-prevalence areas even within nations otherwise succeeding in reducing overall CGF prevalence. By highlighting where the highest-need populations reside, these geospatial estimates can support policy-makers in planning interventions that are adapted locally and in efficiently directing resources towards reducing CGF and its health implications. © 2020, The Author(s)

Mapping subnational HIV mortality in six Latin American countries with incomplete vital registration systems

BackgroundHuman immunodeficiency virus (HIV) remains a public health priority in Latin America. While the burden of HIV is historically concentrated in urban areas and high-risk groups, subnational estimates that cover multiple countries and years are missing. This paucity is partially due to incomplete vital registration (VR) systems and statistical challenges related to estimating mortality rates in areas with low numbers of HIV deaths. In this analysis, we address this gap and provide novel estimates of the HIV mortality rate and the number of HIV deaths by age group, sex, and municipality in Brazil, Colombia, Costa Rica, Ecuador, Guatemala, and Mexico.MethodsWe performed an ecological study using VR data ranging from 2000 to 2017, dependent on individual country data availability. We modeled HIV mortality using a Bayesian spatially explicit mixed-effects regression model that incorporates prior information on VR completeness. We calibrated our results to the Global Burden of Disease Study 2017.ResultsAll countries displayed over a 40-fold difference in HIV mortality between municipalities with the highest and lowest age-standardized HIV mortality rate in the last year of study for men, and over a 20-fold difference for women. Despite decreases in national HIV mortality in all countries-apart from Ecuador-across the period of study, we found broad variation in relative changes in HIV mortality at the municipality level and increasing relative inequality over time in all countries. In all six countries included in this analysis, 50% or more HIV deaths were concentrated in fewer than 10% of municipalities in the latest year of study. In addition, national age patterns reflected shifts in mortality to older age groups-the median age group among decedents ranged from 30 to 45years of age at the municipality level in Brazil, Colombia, and Mexico in 2017.ConclusionsOur subnational estimates of HIV mortality revealed significant spatial variation and diverging local trends in HIV mortality over time and by age. This analysis provides a framework for incorporating data and uncertainty from incomplete VR systems and can help guide more geographically precise public health intervention to support HIV-related care and reduce HIV-related deaths.Peer reviewe