Myosin-X functions in polarized epithelial cells

Myosin-X, an unconventional myosin that has been studied primarily in fibroblast-like cells, has been shown to have important functions in polarized epithelial cell junction formation, regulation of paracellular permeability, and epithelial morphogenesis.Myosin-X (Myo10) is an unconventional myosin that localizes to the tips of filopodia and has critical functions in filopodia. Although Myo10 has been studied primarily in nonpolarized, fibroblast-like cells, Myo10 is expressed in vivo in many epithelia-rich tissues, such as kidney. In this study, we investigate the localization and functions of Myo10 in polarized epithelial cells, using Madin-Darby canine kidney II cells as a model system. Calcium-switch experiments demonstrate that, during junction assembly, green fluorescent protein–Myo10 localizes to lateral membrane cell–cell contacts and to filopodia-like structures imaged by total internal reflection fluorescence on the basal surface. Knockdown of Myo10 leads to delayed recruitment of E-cadherin and ZO-1 to junctions, as well as a delay in tight junction barrier formation, as indicated by a delay in the development of peak transepithelial electrical resistance (TER). Although Myo10 knockdown cells eventually mature into monolayers with normal TER, these monolayers do exhibit increased paracellular permeability to fluorescent dextrans. Importantly, knockdown of Myo10 leads to mitotic spindle misorientation, and in three-dimensional culture, Myo10 knockdown cysts exhibit defects in lumen formation. Together these results reveal that Myo10 functions in polarized epithelial cells in junction formation, regulation of paracellular permeability, and epithelial morphogenesis

Tracking of Salmonella Positive Pigs from Farm to Fork in the Republic of Ireland

In this study, individual pigs from selected herds of known Salmonella serological status were tracked through the slaughter and dressing process. From all tracked animals, caecal contents, rectal faeces, carcasses (before washing and chilling and after chilling) and pork primal cuts were examined for the presence of Salmonella. All samples were screened for Salmonella using real time PCR and all suspect positive samples were confirmed using the ISO 6579 method for Salmonella. To determine the relationship between Salmonella isolates from different parts of the chain , all isolates are being characterised by Pulse Field Gel Electrophoresis (PFGE). The results suggest that the slaughter and dressing operations have a significant effect on the incidence of Salmonella and that even if pigs are presented for slaughter with caecal or rectal carriage of Salmonella then good slaughter practices can prevent carcass contamination. All data generated in the study is being fed into a quantitative risk assessment model for Salmonella in pork

Surface Attachment of Active Antimicrobial Coatings Onto Conventional Plastic Based Laminates and Performance Assessment of These Materials on the Storage Life of Vacuum Packaged Beef Sub-Primals

Two antimicrobial coatings, namely Sodium octanoate and Auranta FV (a commercial antimicrobial composed of bioflavonoids, citric, malic, lactic, and caprylic acids) were used. These two antimicrobials were surface coated onto the inner polyethylene layer of cold plasma treated polyamide films using beef gelatin as a carrier and coating polymer. This packaging material was then used to vacuum pack beef sub-primal cuts and stored at 4 °C. A control was prepared using the non-coated commercial laminate and the same vacuum packaged sub-primal beef cuts. During storage, microbial and quality assessments were carried out. Sodium octanoate treated packages significantly (p \u3c 0.05) reduced microbial counts for all bacteria tested with an increase of 7 and 14 days, respectively compared to control samples. No significant effect on pH was observed with any treatment. The results suggested that these food grade antimicrobials have the potential to be used in antimicrobial active packaging applications for beef products

The influence of acid tolerance response and diet on the survival of Salmonella serotypes through the gastrointestinal tract of pigs

Research findings suggest that acid supplementation of feed and feed characteristics affect the survival of Salmonella in pigs although results are not always consistent. The acid tolerance response (ATR) of Salmonella, induced by mildly acidic conditions, which can occur both within and outside the host, may influence the survival and transmission of Salmonellae in pigs fed certain diets. This study investigates the role of the pH-dependent ATR on relative survival of five Salmonella serotypes in gastrointestinal contents of pigs fed regular or acid-supplemented feeds. Stomach, caecal and colon contents were obtained from pigs fed four different diets (6 pigs/diet, pelleted or meal feed with or without potassium difomwte, KDF)

Factors associated with the weight of individual primal cuts and their inter-relationship in cattle

peer-reviewedInput parameters for decision support tools are comprised of, amongst others, knowledge of the associated factors and the extent of those associations with the animal-level feature of interest. The objective of the present study was to quantify the association between the animal-level factors with primal cut yields in cattle and to understand the extent of the variability in primal cut yields independent of other primal cuts or as carcass weight itself. The data used consisted of the weight of 14 primal carcass cuts (as well as carcass weight, conformation and fat score) on up to 54,250 young cattle slaughtered between 2013 and 2017. Linear mixed models, with contemporary group of herd-sex-season of slaughter as a random effect, were used to quantify the associations between a range of model fixed effects with each primal cut separately. Fixed effects in the model were dam parity, heterosis coefficient, recombination loss, a covariate per breed representing the proportion of Angus, Belgian Blue, Charolais, Jersey, Hereford, Limousin, Simmental, and Holstein-Friesian and a three-way interaction between whether the animal was born in a dairy or beef herd, sex, and age at slaughter, with or without carcass weight as a covariate in the mixed model. The raw correlations among all cuts were all positive varying from 0.33 (between the bavette and the striploin) to 0.93 (between the topside and knuckle). The partial correlation among cuts, following adjustment for differences in carcass weight, varied from -0.36 to 0.74. Age at slaughter, sex, dam parity and breed were all associated (P<0.05) with the primal cut weight. Knowledge of the relationship between the individual primal cuts, and the solutions from the models developed in the study, could prove useful inputs for decision support systems level to increase performance

Integronlike Structures in Campylobacter spp. of Human and Animal Origin

Resistance to antimicrobial agents used to treat severe Campylobacter spp. gastroenteritis is increasing worldwide. We assessed the antimicrobial resistance patterns of Campylobacter spp. isolates of human and animal origin. More than half (n = 32) were resistant to sulphonamide, a feature known to be associated with the presence of integrons. Analysis of these integrons will further our understanding of Campylobacter spp. epidemiology

Cortactin is necessary for E-cadherin–mediated contact formation and actin reorganization

Classical cadherin adhesion molecules are key determinants of cell–cell recognition during development and in post-embryonic life. A decisive step in productive cadherin-based recognition is the conversion of nascent adhesions into stable zones of contact. It is increasingly clear that such contact zone extension entails active cooperation between cadherin adhesion and the force-generating capacity of the actin cytoskeleton. Cortactin has recently emerged as an important regulator of actin dynamics in several forms of cell motility. We now report that cortactin is recruited to cell–cell adhesive contacts in response to homophilic cadherin ligation. Notably, cortactin accumulates preferentially, with Arp2/3, at cell margins where adhesive contacts are being extended. Recruitment of cortactin is accompanied by a ligation-dependent biochemical interaction between cortactin and the cadherin adhesive complex. Inhibition of cortactin activity in cells blocked Arp2/3-dependent actin assembly at cadherin adhesive contacts, significantly reduced cadherin adhesive contact zone extension, and perturbed both cell morphology and junctional accumulation of cadherins in polarized epithelia. Together, our findings identify a necessary role for cortactin in the cadherin–actin cooperation that supports productive contact formation

Genome sequence of an Enterobacter helveticus strain, 1159/04 (= LMG 23733), isolated from fruit powder

We report the draft genome sequence of Enterobacter helveticus strain LMG 23733, isolated from fruit powder. The draft genome assembly for E. helveticus strain LMG 23733 has a size of 4,635,476 bp and a G+C content of 55.9%

The Transforming Rho Family GTPase Wrch-1 Disrupts Epithelial Cell Tight Junctions and Epithelial Morphogenesis

Wrch-1, an atypical and transforming Rho GTPase, regulates cellular activities including proliferation and actin organization, but its functions and effectors remain poorly characterized. We show here that Wrch-1 distributes along the apical and basolateral membranes in MDCK cells and binds the cell polarity protein Par6 in a GTP-dependent manner. Activated Wrch-1 negatively regulates the kinetics of tight junction (TJ) assembly during epithelial cell polarization but has no detectable effect on overall cell polarity in confluent monolayers. It also causes a dramatic cytoskeletal reorganization and multilayering in cells grown in two-dimensional culture and disrupts cystogenesis of cells grown in three-dimensional (3D) culture. Similarly, short hairpin RNA-mediated knockdown of Wrch-1 perturbs cystogenesis in 3D culture, suggesting that tight regulation of Wrch-1 activity is necessary for normal epithelial morphogenesis. A weakly transforming effector domain mutant of activated Wrch-1 that inhibits Par6 binding abrogates the ability of Wrch-1 to disrupt TJ formation, actin organization, and epithelial morphogenesis. We hypothesize that Wrch-1-induced morphological and growth transformation may occur in part through Par6-mediated disruption of TJs and actin organization