Effect of sperm penetration in vitro on completion of first meiosis by bovine oocytes arrested at various stages in culture.

Bovine oocytes cultured for 12-20 h in TC-199 were incubated for 24 h in fertilization medium, Brackett and Oliphant medium with bovine serum albumin (10 mg ml-1), caffeine (5 mmoll-1) and heparin (10 micrograms ml-1), with or without frozen-thawed spermatozoa. High penetration rates (93-96%) and significantly (P < 0.001) higher maturation rates were obtained in oocytes incubated with (93-100%) than without (62-72%) spermatozoa. However, when oocytes at the germinal vesicle stage were cultured for 44 h fertilization medium, maturation of oocytes to metaphase II was reduced. However, all oocytes that were first cultured for 20 h and further for 24 h with spermatozoa were penetrated and 40% of the penetrated oocytes reached metaphase II. All of the remaining oocytes that did not mature arrested at the stages of condensed germinal vesicle (39%) or prometaphase I (22%). These results indicate that oocytes at metaphase I at and after sperm penetration are stimulated by sperm penetration to complete maturation.     

Transcriptome and toxin family analysis of the paralysis tick, Ixodes holocyclus

The Australian paralysis tick (Ixodes holocyclus) secretes neuropathic toxins into saliva that induce host paralysis. Salivary glands and viscera were dissected from fully engorged female I. holocyclus ticks collected from dogs and cats with paralysis symptoms. cDNA from both tissue samples were sequenced using Illumina HiSeq 100?bp pair end read technologies. Unique and non-redundant holocyclotoxin sequences were designated as HT2–HT19, as none were identical to the previously described HT1. Specific binding to rat synaptosomes was determined for synthetic HTs, and their neurotoxic capacity was determined by neonatal mouse assay. They induced a powerful paralysis in neonatal mice, particularly HT4 which produced rapid and strong respiratory distress in all animals tested. This is the first known genomic database developed for the Australian paralysis tick. The database contributed to the identification and subsequent characterization of the holocyclotoxin family that will inform the development of novel anti-paralysis control methods.     

Long-term culture of porcine esophageal epithelial cells without use of feeder layers

Epithelial cells from normal pig esophagus survived in culture for about 4 months undergoing about 12 passages and nearly 40 doublings before showing signs of slow proliferation and senescence. Epithelial cells did not show any attachment and proliferation in serum free media, compared to cells supplemented with 10% serum, where the doubling time was between 48 and 60h. Fibroblasts never became the prominent cell type in these cultures at any given time point. The epithelial cells reacted with antibodies to keratin AE1/AE3, keratin 14 and to involucrin, the differentiating marker.     

An Organic Anode for High Temperature Potassium‐Ion Batteries

The wide applications of rechargeable batteries require state‐of‐the‐art batteries that are sustainable (abundant resource), tolerant to high‐temperature operations, and excellent in delivering high capacity and long‐term cycling life. Due to the scarcity and uneven distribution of lithium, it is urgent to develop alternative rechargeable batteries. Herein, an organic compound, azobenzene‐4,4′‐dicarboxylic acid potassium salts (ADAPTS) is developed, with an azo group as the redox center for high performance potassium‐ion batteries (KIBs). The extended π‐conjugated structure in ADAPTS and surface reactions between ADAPTS and K‐ions enable the stable charge/discharge of K‐ion batteries even at high temperatures up to 60 °C. When operated at 50 °C, ADAPTS anode delivers a reversible capacity of 109 mAh g^?1 at 1C for 400 cycles. A reversible capacity of 77 mAh g^?1 is retained at 2C for 1000 cycles. At 60 °C, the ADAPTS‐based KIBs deliver a high capacity of 113 mAh g^?1 with 81% capacity retention at 2C after 80 cycles. The exceptional electrochemical performance demonstrates that ADAPTS is a promising electrode material for high‐temperature KIBs.     

Effect of self-association on the structural organization of partially folded proteins: inactivated actin

The propensity to associate or aggregate is one of the characteristic properties of many nonnative proteins. The aggregation of proteins is responsible for a number of human diseases and is a significant problem in biotechnology. Despite this, little is currently known about the effect of self-association on the structural properties and conformational stability of partially folded protein molecules. G-actin is shown to form equilibrium unfolding intermediate in the vicinity of 1.5 M guanidinium chloride (GdmCl). Refolding from the GdmCl unfolded state is terminated at the stage of formation of the same intermediate state. An analogous form, known as inactivated actin, can be obtained by heat treatment, or at moderate urea concentration, or by the release of Ca(2+). In all cases actin forms specific associates comprising partially folded protein molecules. The structural properties and conformational stability of inactivated actin were studied over a wide range of protein concentrations, and it was established that the process of self-association is rather specific. We have also shown that inactivated actin, being denatured, is characterized by a relatively rigid microenvironment of aromatic residues and exhibits a considerable limitation in the internal mobility of tryptophans. This means that specific self-association can play an important structure-forming role for the partially folded protein molecules.     

Effects of Exin on the development of microbial infection and ethylene release in plants

Effects of Exin on infection of tomato, potato, and cabbage plants withPseudomonas solanacearum andErwinia carotovora and a fungusSclervtium rolfsii were studied. The treatment of infected plants with Exin caused no significant effect on the development of the disease. Treatment with streptomycin as a standard for comparison completely inhibited the growth of these microorganisms. Pretreatment with Exin for one to eight days before infection inhibited the development of diseases. The numbers of tomato and potato plants damaged among those infected withP. solanacearum were lower by 10 and 35% respectively. In field experiments (350 plants per variant), treatment with Exin decreased the development of wilt caused byS. rolfsii andP. solanacearum and rot caused byE. carotovora. Treatment with Exin activated the release of ethylene for not less than 30 days. Possible mechanisms of the effects of Exin are discussed.     

Determination of the cell tropism of serotype 1 feline infectious peritonitis virus using the spike affinity histochemistry in paraffin‐embedded tissues

Unlike for serotype II feline coronaviruses (FCoV II), the cellular receptor for serotype I FCoV (FCoV I), the most prevalent FCoV serotype, is unknown. To provide a platform for assessing the pattern by which FCoV I attaches to its host receptor(s), HEK293 cell lines that stably express the ectodomains of the spike (S) proteins derived from a FCoV I feline enteric coronavirus strain UU7 (FECV UU7) and a feline infectious peritonitis virus strain UU4 (FIPV UU4) were established. Using the recombinant S proteins as probes to perform S protein affinity histochemistry in paraffin‐embedded tissues, although no tissue or enteric binding of FECV UU7 S protein was detected, it was found that by immunohistochemistry that the tissue distribution of FIPV UU4 S protein‐bound cells correlated with that of FIPV antigen‐positive cells and lesions associated with FIP and that the affinity binding of FIPV UU4 S protein on macrophages was not affected by enzymatic removal of host cell‐surface sialic acid with neuraminidase. These findings suggest that a factor(s) other than sialic acid contribute(s) to the macrophage tropism of FIPV strain UU4. This approach allowed obtaining more information about both virus–host cell interactions and the biological characteristics of the unidentified cellular receptor for FCoV I.

     

The first range-wide assessment of Saddle-billed Stork Ephippiorhynchus senegalensis distribution

The Saddle-billed Stork Ephippiorhynchus senegalensis exemplifies a case in conservation research in which a species is assessed as Least Concern on the IUCN Red List and the resulting consideration of low conservation priority has precluded proper scientific study. As a first step in understanding this stork’s true status, we collated all available data to develop a distribution map and then investigated range-wide patterns of occurrence. The updated map greatly improves on past knowledge of the stork’s distribution and helps to identify regions where range contractions have occurred, particularly in Central Africa and parts of West Africa. We found that the stork’s distribution closely overlaps with protected areas and that there has been an overall increase in surface water (largely manmade water bodies)—a proxy for habitat—across the species’ extent of occurrence in recent decades. While this research represents a valuable contribution to our understanding of the Saddle-billed Stork, it also highlights the need for unbiased empirical data, especially from areas that are poorly surveyed, for developing a science-based conservation status assessment.     

Impact of Smoking and Brain Metastasis on Outcomes of Advanced EGFR Mutation Lung Adenocarcinoma Patients Treated with First Line Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors

ObjectivesThis purpose of this study was to examine clinical-pathologic factors – particularly smoking and brain metastases – in EGFR mutation positive (M+) lung adenocarcinoma (ADC) to determine their impact on survival in patients treated with first line EGFR TKI.MethodsA retrospective review of EGFR mutation reflex testing experience for all ADC diagnosed at a tertiary Asian cancer centre from January 2009 to April 2013. Amongst this cohort, patients with advanced EGFR M+ ADC treated with first line EGFR TKI were identified to determine factors that influence progression free and overall survival.Results444/742 (59.8%) ADC reflex tested for EGFR mutations were EGFR M+. Amongst never-smokers (n=468), EGFR M+ were found in 74.5% of females and 76.3% of males, and amongst ever smokers (n=283), in 53.3% of females and 35.6% of males. Exon 20 mutations were found more commonly amongst heavy smokers (> 50 pack years and > 20 pack years, Pearson’s chi square p=0.044, and p=0.038 respectively). 211 patients treated with palliative first line TKI had a median PFS and OS of 9.2 and 19.6 months respectively. 26% of patients had brain metastasis at diagnosis. This was significantly detrimental to overall survival (HR 1.85, CI 1.09-3.16, p=0.024) on multivariate analysis. There was no evidence that smoking status had a significant impact on survival.ConclusionsThe high prevalence of EGFR M+ in our patient population warrants reflex testing regardless of gender and smoking status. Smoking status and dosage did not impact progression free or overall survival in patients treated with first line EGFR TKI. The presence of brain metastasis at diagnosis negatively impacts overall survival.