bluuebunny/crossref_metadata_2025_split huggingface dataset:
A spontaneous missense mutation in the chromodomain helicase DNA‐binding protein 8 (CHD8) gene: a novel association with congenital myasthenic syndrome
Authors: Lee C. Y. , Petkova M. , Morales‐Gonzalez S. , Gimber N. , Schmoranzer J. , Meisel A. , Böhmerle W. , Stenzel W. , Schuelke M. , Schwarz J. M. 
Published: April 2020

AimsCongenital myasthenic syndromes (CMS) are characterized by muscle weakness, ptosis and episodic apnoea. Mutations affect integral protein components of the neuromuscular junction (NMJ). Here we searched for the genetic basis of CMS in female monozygotic twins.MethodsWe employed whole‐exome sequencing for mutation detection and Sanger sequencing for segregation analysis. Immunohistology was done with antibodies against CHD8, rapsyn, β‐catenin (βCAT) and golgin on fi‐bro‐blasts, human and mouse muscle. We recorded superresolution images of the NMJ using 3D‐structured illumination microscopy.ResultsWe discovered a spontaneous missense mutation in CHD8 [chr14:g.21,884,051G>A, GRCh37.p11 | c.1732C>T, NM_00117062 | p.(R578C)], the gene encoding chromodomain helicase DNA‐binding protein 8. This is the first missense mutation affecting Duplin, the short 110 kDa isoform of CHD8. It is known that CHD8/Duplin negatively regulates βCAT signalling in the WNT pathway and plays a role in chromatin remodelling. Inactivating CHD8 mutations are associated with autism spectrum disorder and intellectual disability in combination with facial dysmorphism, overgrowth and macrocephalus. No muscle‐specific phenotype has been reported to date. Co‐immunostaining with rapsyn on human and mouse muscle revealed a strong presence of CHD8 at the NMJ being located towards the sarcoplasmic side of the rapsyn cluster, where it co‐localizes with βCAT.ConclusionWe hypothesize CHD8 to have a role in the maintenance of the structural integrity and function of the NMJ. Both patients benefited from treatment with 3,4‐diaminopyridine, a reversible blocker of voltage‐gated potassium channels at the nerve terminal that prolongs the action potential and increases acetylcholine release.
East/West Visiting Scholars in Pediatric Anesthesia Program (ViSiPAP): Developing tomorrow’s pediatric anesthesia leaders
Authors: Ferschl Marla B. , Lee Jennifer K. , Lockman Justin L. , Black Stephanie , Chatterjee Debnath , Agarwal Rita , Schwartz Lawrence I. , Fiadjoe John , Heitmiller Eugenie , Mershon Bommy Hong , Deutsch Nina , McCloskey John , Infosino Andrew 
Published: April 2020

AbstractPromoting and retaining junior faculty are major challenges for many medical schools. High clinical workloads often limit time for scholarly projects and academic development, especially in anesthesiology. To address this, we created the East/West Visiting Scholars in Pediatric Anesthesia Program (ViSiPAP). The program's goal is to help “jumpstart” academic careers by providing opportunities for national exposure and recognition through invited lectures and collaborative opportunities. East/West ViSiPAP benefits the participating scholars, the home and hosting anesthesia departments, and pediatric anesthesia fellowship training programs. By fostering a sense of well‐being and inclusion in the pediatric anesthesia community, East/West ViSiPAP has the potential to increase job satisfaction, help faculty attain promotion, and reduce attrition. Faculty and trainees are exposed to new expertise and role models. Moreover, ViSiPAP provides opportunities for women and underrepresented in medicine faculty. This program can help develop today's junior faculty into tomorrow's leaders in pediatric anesthesia. We advocate for expanding the concept of ViSiPAP to other institutions in academic medicine.
Endobronchial coils for emphysema: Dual mechanism of action on lobar residual volume reduction
Authors: Hartman Jorine E. , Shah Pallav L. , Sciurba Frank , Herth Felix J.F. , Slebos Dirk‐Jan ,   RENEW Study Group
Published: April 2020

Residual lobar volume reduction in treated lobes measured by QCT was found to be the driving mechanism of action of endobronchial coils leading to positive clinical outcomes. However, the improvement in exercise capacity and quality of life seems to be affected by the presence of cardiac disease.See related Editorial
Conservation science and technology identity instrument: Empirically measuring STEM identities in informal science learning programs
Authors: Rodriguez Laura S. , Morzillo Anita , Volin John C. , Campbell Todd 
Published: April 2020

AbstractOur research centered on developing the Conservation Science and Technology Identity (CSTI) instruments as an empirical way to measure STEM identities and the intersection of identity constructs such as competence, performance, recognition, and ways of seeing and being. The surveys were used in a large funded multi‐year project for teens and adults learning geospatial technologies and conservation science to use in intergenerational community conservation projects. We investigated whether an informal STEM learning program was developing new STEM identities or advancing well‐developed identities. The instruments’ content validity was determined through a vetting process from national STEM identity research experts. Reliability was estimated with Cronbach coefficient alpha. Mann‐Whitney and Wilcoxon Signed Rank tests were used to determine participants’ STEM identities and the workshop’s effect on specific identity constructs. We found teens and adults had historically similar STEM identities, with stronger conservation science than technology. Both science and technology competences, as well as technology ways of seeing and being, significantly increased, suggesting CSTI can be a valuable instrument in empirically assessing STEM identities.
Increase of CD26/Dipeptidyl Peptidase IV Expression on Human Gingival Fibroblasts upon Stimulation with Cytokines and Bacterial Components
Authors: Nemoto Eiji , Sugawara Shunji , Takada Haruhiko , Shoji Shigeru , Horiuch Hiroshi 
Published: April 2020

ABSTRACTCD26/dipeptidyl peptidase IV (DPPIV) is a cell surface ectoenzyme which participates in immune and inflammatory reactions. We found that CD26 was only partially expressed on human fibroblasts from periodontal tissues, whereas fibroblasts from lung and skin expressed CD26 constitutively as revealed by flow cytometry. We examined the possible upregulation of CD26 expression on human gingival fibroblasts in response to various stimulants. Interleukin-1α (IL-1α); tumor necrosis factor alpha; gamma interferon; lipopolysaccharide fromPorphyromonas gingivalis,Prevotella intermedia, andEscherichia coli; andPrevotellaglycoprotein augmented CD26 expression on gingival fibroblasts. Among the stimulants, IL-1α exhibited the most potent activity. Enzymatic activity of CD26 induced by IL-1α on fibroblasts was determined colorimetrically in terms of Gly-Pro hydrolysis of a synthetic chromogenic substrate, Gly-Prop-nitroanilide. Among various inhibitors tested, diprotin A and phenylmethylsulfonyl fluoride inhibited the enzymatic activity, suggesting that the enzyme induced by IL-1α was DPPIV. The upregulation of CD26 mRNA expression upon stimulation with IL-1α was also revealed by a quantitative reverse transcription-PCR assay. In the kinetic experiment, 48 h and several days were required for maximum CD26 mRNA accumulation and CD26 molecule expression on the cell surface, respectively. The addition of cycloheximide at 2 h before IL-1α stimulation almost completely inhibited the accumulation of CD26 mRNA. These results suggested that induction of CD26 on human gingival fibroblasts is regulated at the transcriptional level and is also dependent on a de novo-synthesized protein factor(s).
Expression of Recombinant EnterotoxigenicEscherichia coliColonization Factor Antigen I bySalmonella typhimuriumElicits a Biphasic T Helper Cell Response
Authors: Pascual David W. , Hone David M. , Hall Stacy , van Ginkel Frederik W. , Yamamoto Masafumi , Walters Nancy , Fujihashi Kohataro , Powell Robert J. , Wu Shaoguang , Vancott John L. , Kiyono Hiroshi , McGhee Jerry R. 
Published: April 2020

ABSTRACTProtective immunity to enterotoxigenicEscherichia coli(ETEC) is antibody (Ab) dependent; however, oral immunization with purified ETEC fimbriae fails to elicit protective immunity as a consequence of antigenic alteration by the gastrointestinal (GI) tract. Unless unaltered ETEC fimbriae can reach the inductive lymphoid tissues of the GI tract, immunity to ETEC cannot be induced. To produce immunity, live vectors, such asSalmonella typhimurium, can effectively target passenger antigens to the inductive lymphoid tissues of the GI tract. By convention, oral immunizations withSalmonellavectors induce CD4+T helper (Th) cell responses by gamma interferon (IFN-γ)-dominated pathways both to the vector and passenger antigen, resulting in serum immunoglobulin G2a (IgG2a) and modest mucosal IgA Ab responses. In the present study, mice orally immunized with aSalmonellavector engineered to stably express ETEC colonization factor antigen I (CFA/I) showed initially elevated serum IgG1 and mucosal IgA anti-CFA/I Ab responses. As expected, mice orally immunized with anE. coli-CFA/I construct elicited poor anti-CFA/I Ab responses. In fact, the addition of cholera toxin during oralE. coli-CFA/I immunization failed to greatly enhance mucosal IgA Ab responses. Seven days after immunization with theSalmonella-CFA/I construct, cytokine-specific ELISPOT showed induction of predominant Th2-type responses in both mucosal and systemic immune compartments supporting the early IgG1 and IgA anti-CFA/I Abs. By 4 weeks, the Th cell response became Th1 cell dominant from the earlier Th2-type responses, as evidenced by increased mucosal and systemic IFN-γ-producing T cells and a concomitant elevation of serum IgG2a Ab responses. This biphasic response offers an alternative strategy for directingSalmonellavector-induced host immunity along a Th2 cell-dependent pathway, allowing for early promotion of mucosal and systemic Abs.
Cytokine Profiles of AIDS Patients Are Similar to Those of Mice with DisseminatedCryptococcus neoformansInfection
Authors: Lortholary Olivier , Improvisi Luce , Rayhane Naima , Gray Francoise , Fitting Catherine , Cavaillon Jean Marc , Dromer Francoise 
Published: April 2020

ABSTRACTCryptococcosis is an hematogenously disseminated meningoencephalitis during which the relationship between the disease severity and the immune response remains unclear. We thus analyzed, by enzyme-linked immunosorbent assay, proinflammatory (tumor necrosis factor alpha [TNF-α] and interleukin-6 [IL-6]) and anti-inflammatory (IL-10) cytokine levels in plasma at the time of diagnosis in 51 AIDS patients with culture-proven cryptococcosis. We used a murine model to determine the correlation between cytokine levels and fungal burden in blood and tissues and the kinetics of the immune response and of the formation of cerebral lesions. In AIDS patients, plasma TNF-α and IL-10, but not IL-6, levels were significantly higher in the case of fungemia or disseminated infection than in their absence, whereas the presence of meningitis had no influence on these levels. In mice, none of these cytokines were detected within the first day after inoculation. Later on, TNF-α and IL-10, but not IL-6, levels in plasma correlated significantly with the fungal burden in the blood and spleen but not the brain. In the brain, cytokine levels were low compared to those in other compartments, and tissue lesions and a degree of infection similar to those observed in humans were seen, further suggesting the relevance of this experimental model. Thus, AIDS patients with cryptococcosis produce an immune response that reflects the dissemination but not the meningeal involvement. This murine model of disseminated cryptococcosis can be used to investigate the pathophysiology of cryptococcosis and new therapeutic approaches.
Immunoglobulin Concentrations and Antigen-Specific Antibody Levels in Cervicovaginal Lavages of Rhesus Macaques Are Influenced by the Stage of the Menstrual Cycle
Authors: Lü F. Xusheng , Ma Zhongmin , Rourke Tracy , Srinivasan Seema , McChesney Michael , Miller Christopher J. 
Published: April 2020

ABSTRACTThe levels of antigen-specific antibodies (Abs) and immunoglobulins in the cervical mucus of women vary with the menstrual cycle; the highest levels occur during menses, and the lowest occur during the periovulatory period. The rhesus macaque is a widely used animal model of female genital tract immunity. We sought to determine whether rhesus macaques have a cyclical pattern of changing cervicovaginal Ab and immunoglobulin levels that is similar to that of the human female. This study examined the relationship of the stages of the menstrual cycle to genital mucosal and systemic immunoglobulin concentrations and Ab levels in rhesus macaques. In all seven rhesus macaques studied, the immunoglobulins G and A and some antibodies in cervicovaginal lavages varied with the stages of the menstrual cycle, and in many cases this variation reached the level of statistical significance. In a pattern similar to that of women, the highest levels of Abs and immunoglobulins occurred during menses, and the lowest levels occurred around the time of ovulation. However, the Ab and immunoglobulin levels in serum and rectal lavages did not change with the menstrual cycle stage. The results of this study are consistent with the hypothesis that the ovarian hormones that drive the menstrual cycle influence genital tract immunity in female primates.
Characterization of the
angR
Gene of
Vibrio anguillarum
: Essential Role in Virulence
Authors: Wertheimer A. M. , Verweij W. , Chen Q. , Crosa L. M. , Nagasawa M. , Tolmasky M. E. , Actis L. A. , Crosa J. H. 
Published: April 2020

ABSTRACT

The ability to utilize the iron bound by high-affinity iron-binding proteins in the vertebrate host is an important virulence factor for the marine fish pathogen
Vibrio anguillarum
. Virulence in septicemic infections is due to the presence of a highly efficient plasmid-encoded iron transport system. AngR, a 110-kDa protein component of this system, appears to play a role in both regulation of the expression of the iron transport genes
fatDCBA
and the production of the siderophore anguibactin. Therefore, study of the expression of the
angR
gene and the properties of its product, the AngR protein, may contribute to the understanding of the mechanisms of virulence of this pathogen. In this work, we present genetic and molecular evidence from transposition mutagenesis experiments and RNA analysis that
angR
, which maps immediately downstream of the
fatA
gene, is part of a polycistronic transcript that also includes the iron transport genes
fatDCBA
and
angT
, a gene located downstream of
angR
which showed domain homology to certain thioesterases involved in nonribosomal peptide synthesis of siderophores and antibiotics. In order to dissect the specific domains of AngR associated with regulation of iron transport gene expression, anguibactin production, and virulence, we also generated a panel of site-directed
angR
mutants, as well as deletion derivatives. Both virulence and anguibactin production were dramatically affected by each one of the
angR
modifications. In contrast to the need for an intact AngR molecule for anguibactin production and virulence, the regulation of iron transport gene expression does not require the entire AngR molecule, since truncation of the carboxy terminus carrying the nonribosomal peptide synthetase cores, as well as the site-directed mutations, resulted in derivatives that retained their ability to regulate gene expression which was only abolished after truncation of amino-terminal sequences containing helix-turn-helix and leucine zipper motifs and a specialized heterocyclization and condensation domain found in certain nonribosomal peptide synthetases. The evidence, while not rigorously eliminating the possibility that a separate regulatory polypeptide exists and is encoded somewhere within the 5′-end region of the
angR
gene, strongly supports the idea that AngR is a bifunctional protein and that it plays an essential role in the virulence mechanisms of
V. anguillarum
. We also show in this study that the
angT
gene, found downstream of
angR
, intervenes in the mechanism of anguibactin production but is not essential for virulence or iron transport gene expression.
ThepspCGene ofStreptococcus pneumoniaeEncodes a Polymorphic Protein, PspC, Which Elicits Cross-Reactive Antibodies to PspA and Provides Immunity to Pneumococcal Bacteremia
Authors: Brooks-Walter Alexis , Briles David E. , Hollingshead Susan K. 
Published: April 2020

ABSTRACTPspC is one of three designations for a pneumococcal surface protein whose gene is present in approximately 75% of allStreptococcus pneumoniaestrains. Under the name SpsA, the protein has been shown to bind secretory immunoglobulin A (S. Hammerschmidt, S. R. Talay, P. Brandtzaeg, and G. S. Chhatwal, Mol. Microbiol. 25:1113–1124, 1997). Under the name CbpA, the protein has been shown to interact with human epithelial and endothelial cells (C. Rosenow et al., Mol. Microbiol. 25:819–829, 1997). The gene is paralogous to thepspAgene inS. pneumoniaeand was thus calledpspC(A. Brooks-Walter, R. C. Tart, D. E. Briles, and S. K. Hollingshead, Abstracts of the 97th General Meeting of the American Society for Microbiology 1997). Sequence comparisons of five published and seven new alleles reveal that this gene has a mosaic structure, and modular domains have contributed to gene diversity during evolution. Two major clades exist: clade A alleles are larger and contain an extra module that is shared with manypspAalleles; clade B alleles are smaller and lack thispspA-like domain. All alleles have a proline-rich domain and a choline-binding repeat domain that show 0% divergence from similar domains in the PspA protein. Immunization of a rabbit with a recombinant clade B PspC molecule produced antiserum that cross-reacted with both PspC and PspA from 15 pneumococcal isolates. The cross-reactive antibodies afforded cross-protection in a mouse model system. Mice immunized with PspC were protected against challenge with a strain that expressed PspA but not PspC. The PspA- and PspC-cross-reactive antibodies were directed to the proline-rich domain present in both molecules.
Immunopathologic Alterations in Murine Models of Sepsis of Increasing Severity
Authors: Ebong Samuel , Call Douglas , Nemzek Jean , Bolgos Gerald , Newcomb David , Remick Daniel 
Published: April 2020

ABSTRACTWe investigated inflammatory and physiologic parameters in sepsis models of increasing lethality induced by cecal ligation and puncture (CLP). Mice received imipenem for antibiotic therapy, and groups were sacrificed at 2, 4, 8, 12, 16, 20, and 24 h after CLP. The severity of sepsis increased with needle puncture size (lethality with 18-gauge puncture [18G], 100%; 21G, 50%; 25G, 5%; sham treatment, 0%). While the temperature (at 12 h) and the activity and diurnal rhythm (at day 4) of the 25G-treated CLP group recovered to normal, the 21G and 18G treatment groups exhibited severe hypothermia along with decreased activities. A direct correlation was also observed between the severity of sepsis and cytokine (interleukin 1β [IL-1β], tumor necrosis factor [TNF], IL-6, and IL-10) concentrations in both the peritoneum and the plasma. There were substantially higher cytokine levels in the more severe CLP models than in the sham-treated one. Peritoneal and plasma TNF levels were always less than 40 pg/ml in all models. None of the cytokines in the septic mice peaked within the first hour, which is in contrast to the results of most endotoxin models. Chemokine (KC and macrophage inflammatory protein 2) profiles also correlated with the severity of sepsis. Except for the chemokines, levels of inflammatory mediators were always higher at the site of inflammation (peritoneum) than in the circulation. Our study demonstrated that sepsis of increasing severity induced increased cytokine levels both within the local environment (peritoneum) and systemically (plasma), which in turn correlated with morbidity and mortality.
Peptide Localization and Gene Structure of Cryptdin 4, a Differentially Expressed Mouse Paneth Cell α-Defensin
Authors: Ouellette Andre J. , Darmoul Dalila , Tran Dat , Huttner Kenneth M. , Yuan Jun , Selsted Michael E. 
Published: April 2020

ABSTRACTPaneth cells in crypts of the small intestine express antimicrobial peptides, including α-defensins, termed cryptdins in mice. Of the known Paneth cell α-defensins, the cryptdin 4 gene is unique, because it is inactive in the duodenum and expressed at maximal levels in the distal small bowel (D. Darmoul and A. J. Ouellette, Am. J. Physiol. 271:G68–G74, 1996). With a cryptdin 4-specific antibody, immunohistochemical staining of ileal Paneth cells was strong and specific for cytoplasmic granules, demonstrating that this microbicidal peptide is a secretory product of Paneth cells in the distal small intestine. Consistent with the pattern of cryptdin 4 mRNA distribution along the length of the gut, the cryptdin 4 peptide was not detected in duodenum. Structurally, the cryptdin 4 gene resembles other Paneth cell α-defensin genes. Its two exons, transcriptional start site, intron, splice sites, and 3′ flanking sequences are characteristic of the highly conserved mouse α-defensin genes. However, in the region upstream of the transcriptional initiation site, the cryptdin 4 gene contains a repeated 130-bp element that is unique to this α-defensin gene. Every independent cryptdin 4 genomic clone examined carries the repeated element, which contains putative recognition sequences for TF-IID-EIIA, cMyc-RS-1, and IgHC.2/CuE1.1; the repeat proximal to the start of transcription replaces DNA at the corresponding position in other mouse α-defensin genes. We speculate that this unique duplicated element may have acis-acting regulatory role in the positional specificity of cryptdin 4 gene expression.
Analysis of Antibody Response to Invariable Regions of VlsE, the Variable Surface Antigen of
Borrelia burgdorferi
Authors: Liang Fang Ting , Philipp Mario T. 
Published: April 2020

ABSTRACT

VlsE, the variable surface antigen of
Borrelia burgdorferi
, consists of two invariable domains at the amino and carboxyl termini and one central variable domain. The latter contains six invariable regions, IR
1
to IR
6
, and six variable regions. In the present study, the antigenicity of all of the invariable regions in
B. burgdorferi
-infected monkeys, humans, and mice was assessed by peptide-based enzyme-linked immunosorbent assays. Only one invariable region, IR
6
, was antigenic in all animals of the three host species. IR
2
and IR
4
were also antigenic in mice.
Delivery of a MalE CD4+-T-Cell Epitope into the Major Histocompatibility Complex Class II Antigen Presentation Pathway byBordetella pertussisAdenylate Cyclase
Authors: Loucká Jir̆ina , Schlecht Géraldine , Vodolánová Jana , Leclerc Claude , S̆ebo Peter 
Published: April 2020

ABSTRACTRecombinant adenylate cyclase toxoids are shown to deliver inserted foreign CD4+-T-cell epitopes into the major histocompatibility complex class II presentation pathway, inducing a specific CD4+-T-cell response in vivo and yielding in vitro stimulation of specific CD4+T cells at a 100-times-higher molar efficiency than the free peptide containing the epitope.
Novel Laminin-Binding Protein of
Streptococcus pyogenes
, Lbp, Is Involved in Adhesion to Epithelial Cells
Authors: Terao Yutaka , Kawabata Shigetada , Kunitomo Eiji , Nakagawa Ichiro , Hamada Shigeyuki 
Published: April 2020

ABSTRACT

The
lbp
gene, which encodes a laminin-binding protein (Lbp) of
Streptococcus pyogenes
, was found in all
S. pyogenes
M types. An Lbp-deficient mutant showed a significantly lower efficiency of adhesion to HEp-2 cells than did the wild-type strain. These results indicate that Lbp is one of the important
S. pyogenes
adhesins.