Lipid transfer proteins (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens

Four homogeneous proteins (Cw18, Cw20, Cw21, Cw22) were isolated from etiolated barley leaves by extraction of the insoluble pellet from a Tris-HCl (pH 7.5) homogenate with 1.5 M LiCl and fractionation by reverse-phase high-performance liquid chromatography. All 4 proteins inhibited growth of the pathogen Clavibacter michiganensis subsp. sepedonicus (EC50S = 1−3 × 10−7 M) and had closely related N-terminal amino acid sequences. The complete amino acid sequences of proteins Cw18 and Cw21 were determined and found to be homologous to previously described, non-specific lipid transfer proteins from plants (32–62% identical positions). The proteins also inhibited growth of the bacterial pathogen Pseudomonas solanacearum (EC50s = 3–6 × 10−7 M) and the fungus Furium solani (EC50s = 3–20 × 10−6 M). A homologous protein from maize leaves (Cw41) was purified in a similar manner and also found to have inhibitory properties, A synergistic effect against the fungus was observed when protein Cw21, was combined with thionins. A defense role for non-specific lipid transfer proteins from plants is proposed

A mixed treatment comparison meta-analysis of metaphylaxis treatments for bovine respiratory disease in beef cattle

Citation: Abell, K. M., Theurer, M. E., Larson, R. L., White, B. J., & Apley, M. (2017). A mixed treatment comparison meta-analysis of metaphylaxis treatments for bovine respiratory disease in beef cattle. Journal of Animal Science, 95(2), 626-635. doi:10.2527/jas2016.1062The objective of this project was to evaluate the effects of antimicrobials approved for parenteral metaphylactic use in feeder and stocker calves on morbidity and mortality for bovine respiratory disease with the use of a mixed treatment comparison meta-analysis. An initial literature review was conducted in April 2016 through Pubmed, Agricola, and CAB (Commonwealth Agricultural Bureau) for randomized controlled trials for metaphylaxis antimicrobial administered parentally to incoming feedlot or stocker calves within 48 h of arrival. The final list of publications included 29 studies, with a total of 37 trials. There were 8 different metaphylactic antimicrobials. Final event outcomes were categorized into bovine respiratory disease (BRD) morbidity cumulative incidence d 1 to <= 60 of the feeding period, BRD morbidity cumulative incidence d 1 to closeout of the feeding period, BRD mortality cumulative incidence d 1 to closeout of the feeding period, and BRD retreatment cumulative incidence morbidity d 1 to closeout of the feeding period. Network meta-analysis combined direct and indirect evidence for all the event outcomes to determine mean odds ratio (OR) with 95% credibility intervals (CrIs) for all metaphylactic antimicrobial comparisons. The "upper tier" treatment arms for morbidity d 1 to <= 60 included tulathromycin, gamithromycin, and tilmicosin. For BRD mortality cumulative incidence d 1 to closeout and BRD retreatment morbidity d 1 to closeout, classifying the treatment arms into tiers was not possible due to overlapping 95% CrIs. The results of this project accurately identified differences between metaphylactic antimicrobials, and metaphylactic antimicrobial options appear to offer different outcomes on BRD morbidity and mortality odds in feedlot cattle

Evidence against Zika virus infection of pets and peri-domestic animals in Latin America and Africa

Decades after its discovery in East Africa, Zika virus (ZIKV) emerged in Brazil in 2013 and infected millions of people during intense urban transmission. Whether vertebrates other than humans are involved in ZIKV transmission cycles remained unclear. Here, we investigate the role of different animals as ZIKV reservoirs by testing 1723 sera of pets, peri-domestic animals and African non-human primates (NHP) sampled during 2013–2018 in Brazil and 2006–2016 in Côte d'Ivoire. Exhaustive neutralization testing substantiated co-circulation of multiple flaviviruses and failed to confirm ZIKV infection in pets or peri-domestic animals in Côte d'Ivoire (n=259) and Brazil (n=1416). In contrast, ZIKV seroprevalence was 22.2% (2/9, 95% CI, 2.8–60.1) in West African chimpanzees (Pan troglodytes verus) and 11.1% (1/9, 95% CI, 0.3–48.3) in king colobus (Colobus polycomos). Our results indicate that while NHP may represent ZIKV reservoirs in Africa, pets or peri-domestic animals likely do not play a role in ZIKV transmission cycles.Peer Reviewe

Kupffer cell-monocyte communication is essential for initiating murine liver progenitor cell-mediated liver regeneration

Liver progenitor cells (LPCs) are necessary for repair in chronic liver disease because the remaining hepatocytes cannot replicate. However, LPC numbers also correlate with disease severity and hepatocellular carcinoma risk. Thus, the progenitor cell response in diseased liver may be regulated to optimize liver regeneration and minimize the likelihood of tumorigenesis. How this is achieved is currently unknown. Human and mouse diseased liver contain two subpopulations of macrophages with different ontogenetic origins: prenatal yolk sac-derived Kupffer cells and peripheral blood monocyte-derived macrophages. We examined the individual role(s) of Kupffer cells and monocyte-derived macrophages in the induction of LPC proliferation using clodronate liposome deletion of Kupffer cells and adoptive transfer of monocytes, respectively, in the choline-deficient, ethionine-supplemented diet model of liver injury and regeneration. Clodronate liposome treatment reduced initial liver monocyte numbers together with the induction of injury and LPC proliferation. Adoptive transfer of monocytes increased the induction of liver injury, LPC proliferation, and tumor necrosis factor-a production. Conclusion: Kupffer cells control the initial accumulation of monocyte-derived macrophages. These infiltrating monocytes are in turn responsible for the induction of liver injury, the increase in tumor necrosis factor-a, and the subsequent proliferation of LPCs

Empathy Enhancement Based on a Semiotics Training Program: A Longitudinal Study in Peruvian Medical Students

Background: Empathy, as a core element of medical professionalism, is part of leadership in medicine. This attribute, predominantly cognitive, involves understanding and communication capacity. Empathy can be enhanced with courses on medical semiotics. It appears adequate to apply this enhancement in the early stages of professional training. Based on this, this study was performed with the purpose of demonstrating the positive effect that an academic course on medical semiotics has on the development of empathy in medical students. Methods: A quasi-experimental study was conducted in one School of Medicine in Peru, where medical students had to attend a 17-week course on medical semiotics as part of their regular training. The sample, composed by 269 students, included two cohorts of third-year medical students. As main measures, the Jefferson Scales of Empathy (JSE), inter-professional collaboration (JSAPNC), and lifelong learning (JeffSPLL), were used. In addition, students’ scores evaluating theoretical and practical aspects of the course were collected once the course was finished. Pre- and posttests were administered in week 1 and in week 17. Analyses compared measures in both moments and in time. Inter-professional collaboration and lifelong learning scores and empathy scores were used as discriminant and convergent validity measures of students’ course scores, respectively. Results: Gender differences on empathy appeared, but only at the beginning. In the entire sample, empathy enhancement was confirmed in time (p < 0.001), with a large effect size (r = 0.45). This effect was also observed in both gender groups, separately. On the contrary, no changes appeared in inter-professional collaboration and in lifelong learning abilities in time. In addition, a positive correlation was observed among empathy, inter-professional collaboration and lifelong learning abilities at the beginning and at the end, confirming that the improvement observed was specific for empathy and explained by the educational intervention assessed. Conclusion: These findings bring empiric evidence supporting the positive effect that training in medical semiotics has on empathy. In addition, these findings highlight some gender differences in the development of empathy in medical students.Fundacion Rioja Salud, in SpainUniversidad Privada Antenor Orrego, in Per

An efficient and scalable pipeline for epitope tagging in mammalian stem cells using Cas9 ribonucleoprotein

AbstractCRISPR/Cas9 can be used for precise genetic knock-in of epitope tags into endogenous genes, simplifying experimental analysis of protein function. However, Cas9-assisted epitope tagging in primary mammalian cell cultures is often inefficient and reliant on plasmid-based selection strategies. Here we demonstrate improved knock-in efficiencies of diverse tags (V5, 3XFLAG, Myc, HA) using co-delivery of Cas9 protein pre-complexed with two-part synthetic modified RNAs (annealed crRNA:tracrRNA) and single-stranded oligodeoxynucleotide (ssODN) repair templates. Knock-in efficiencies of ~5-30%, were achieved without selection in embryonic stem (ES) cells, neural stem (NS) cells, and brain tumour-derived stem cells. Biallelic-tagged clonal lines were readily derived and used to define Olig2 chromatin-bound interacting partners. Using our novel web-based design tool, we established a 96-well format pipeline that enabled V5-tagging of sixty different transcription factors. This efficient, selection-free and scalable epitope tagging pipeline enables systematic surveys of protein expression levels, subcellular localization, and interactors across diverse mammalian stem cells.</jats:p

The defensive role of nonspecific lipid-transfer proteins in plants

Plant nonspecific lipid-transfer proteins stimulate the transfer of a broad range of lipids between membranes in vitro. In view of their ability to inhibit bacterial and fungal pathogens, their distribution at high concentrations over exposed surfaces and in the vascular system, and the response of Ltp-gene expression to infection with pathogens, they are now thought to be active plant-defense proteins