The surface protein internalin A (InlA) contributes to the invasion of human intestinal epithelial cells by Listeria monocytogenes. Screening of L. monocytogenes strains isolated from human clinical cases (n ؍ 46), foods (n ؍ 118), and healthy animals (n ؍ 58) in the United States revealed mutations in inlA leading to premature stop codons (PMSCs) in L. monocytogenes ribotypes DUP-1052A and DUP-16635A (PMSC mutation type 1), DUP-1025A and DUP-1031A (PMSC mutation type 2), and DUP-1046B and DUP-1062A (PMSC mutation type 3). While all DUP-1046B, DUP-1062A, DUP-16635A, and DUP-1031A isolates (n ؍ 76) contained inlA PMSCs, ribotypes DUP-1052A and DUP-1025A (n ؍ 72) contained isolates with and without inlA PMSCs. Western immunoblotting showed that all three inlA PMSCs result in the production of truncated and secreted InlA. Searches of the Pathogen Tracker database, which contains subtype and source information for more than 5,000 L. monocytogenes isolates, revealed that the six ribotypes shown to contain isolates with inlA PMSCs were overall more commonly isolated from foods than from human listeriosis cases. L. monocytogenes strains carrying inlA PMSCs also showed significantly (P ؍ 0.0004) reduced invasion of Caco-2 cells compared to isolates with homologous 3 inlA sequences without PMSCs. Invasion assays with an isogenic PMSC mutant further supported the observation that inlA PMSCs lead to reduced invasion of Caco-2 cells. Our data show that specific L. monocytogenes subtypes which are common among U.S. food isolates but rare among human listeriosis isolates carry inlA mutations that are associated with, and possibly at least partially responsible for, an attenuated invasion phenotype.
Based on a Freedom of Information request with data from 75% of all English children's services departments covering over ½ million children, this paper shows that 22.5% of children born in the 2009-10 financial year were referred to children's social care before their fifth birthday. Three-quarters of them were at some point assessed; almost two-thirds found to be in need; and a quarter formally investigated. These findings show the full extent of children's involvement in children's social care before the age of five. One in every 9 children born in 2009-10 was suspected by social workers of being abused and this high level of involvement is only justifiable if it is demonstrably reducing harm and promoting well-being of children -an outcome which is contested. Early Help's introduction was associated with high proportions of children being referred and assessed and rapidly increasing numbers of investigations thus questioning its ability to prevent entry to the child protection system. The paper calls for a change from the current emphasis on individualised and investigative approaches to child protection in order to provide an effective and humane response to children, the majority of whom live in families affected by high levels of deprivation and poverty. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 IntroductionDespite statistics on safeguarding being collected in England for many years, numerous inquiries, and decades of research there has never been information published at a national level to show how many children are involved in the system over their lifetime. This paper reports on a freedom of information (FoI) request that gathered data from 75% of all local authorities (LAs) in England covering over half a million children, 80% of all those born in the 2009-10 financial year. It provides, for the first time, information on how many children were referred to children's services before reaching the age of five. It also shows how many were formally assessed; suspected of abuse; subject to a child protection investigation; on a child protection plan; and entered care. More than one in five of all children were referred and many progressed deeper into the formal child protection system. Given the shame and fear that accusations of abuse can cause (Gibson 2013) this high level of involvement is only justifiable if it is demonstrably reducing harm and promoting well-being of children -a goal which is certainly contested (Edwards et al., 2015; Featherstone, White and Morris, 2014a; Frost and Parton, 2009; Gilbert et al., 2012).The FoI request shows that this information on numbers of children involved in the safeguarding system over 5 years is easily produced. The fact that such important information has, until now, been unavailable is an indication of how research agendas and policy making in many cases have been f...
Over 30 polymorphisms in the KIT Proto-Oncogene Receptor Tyrosine Kinase (KIT) gene have been implicated in white spotting patterns ranging from small areas to full dermal depigmentation in the horse. We performed a candidate-gene exon sequencing approach on KIT and MITF, 2 known causatives of white spotting patterns, within 2 families of horses of unknown white spotting. Family 1 (Fam1, N = 5) consisted of a Quarter Horse stallion and 4 offspring with white spotting pattern ranging from legs, lower ventral, and head regions with jagged borders, to almost complete white. The second family (Fam2, N = 7) consisted of 6 half-sibling American Paint Horse/Quarter Horse and their dam, demonstrating unpigmented limbs with belly spots and an extensive white patterning on the face. This approach resulted in 2 variants significantly associated with familial phenotypes, where Fam1 variant is an indel leading to a frameshift mutation, and Fam2 a non-synonymous SNP. We validated the variants within an unrelated population of horses (Fam2 variant, P = 0.00271944) as well as for protein functional impact with ExPASy, Protter, Phyre2, SMART, PROVEAN, SIFT, and I-TASSER, confirming the reported associations. Fam1 associated variant, deemed W31, alters the protein sequence, leading to an early stop codon truncating the normal amino acid sequence from 972 to just 115 amino acids. Fam2 associated variant, deemed W32, may have a subtle impact on receptor function or could be in linkage with a non-coding or regulatory change creating the mild spotting pattern observed in this family.
White spotting phenotypes in horses are highly valued in some breeds. They are quite variable and may range from the common white markings up to completely white horses. EDNRB, KIT, MITF, PAX3 and TRPM1 represent known candidate genes for white spotting phenotypes in horses. For the present study, we investigated an American Paint Horse family segregating a phenotype involving white spotting and blue eyes. Six of eight horses with the white-spotting phenotype were deaf. We obtained whole-genome sequence data from an affected horse and specifically searched for structural variants in the known candidate genes. This analysis revealed a heterozygous~63-kb deletion spanning exons 6-9 of the MITF gene (chr16:21 503 211-21 566 617). We confirmed the breakpoints of the deletion by PCR and Sanger sequencing. PCR-based genotyping revealed that all eight available affected horses from the family carried the deletion. The finding of an MITF variant fits well with the syndromic phenotype involving both depigmentation and an increased risk for deafness and corresponds to human Waardenburg syndrome type 2A. Our findings will enable more precise genetic testing for depigmentation phenotypes in horses.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.