bluuebunny/crossref_metadata_2025 huggingface dataset:
The Soluble Interleukin-6 (IL-6) Receptor/IL-6 Fusion Protein Enhances In Vitro Maintenance and Proliferation of Human CD34+CD38−/low Cells Capable of Repopulating Severe Combined Immunodeficiency Mice
Authors: Kollet Orit , Aviram Ronit , Chebath Judith , ben-Hur Herzl , Nagler Arnon , Shultz Leonard , Revel Michel , Lapidot Tsvee
Published: October 2019

In vitro maintenance and proliferation of human hematopoietic stem cells is crucial for many clinical applications. Early hematopoietic cells express low levels of FLT-3 and c-kit receptors, as well as the interleukin-6 (IL-6) receptor signal transducing element, gp130, but do not express IL-6 receptor itself. Therefore, we have attempted to maintain human cord blood or bone marrow CD34+ cells ex vivo in serum-free cultures containing stem cell factor (SCF) and FLT-3 ligand (FL) alone or together with a new recombinant molecule of soluble IL-6 receptor fused to IL-6 (IL6RIL6 chimera). The effect of IL6RIL6 chimera on the proliferation and differentiation of CD34+ cells was compared with that of each chimera component added separately. The engraftment potential of in vitro-cultured cells was determined using our recently established functional in vivo assay for primitive human severe combined immunodeficiency (SCID)-repopulating cells (SRC). We report here that IL6RIL6 chimera induced significantly higher levels of progenitors and SRC compared with SCF + FL alone or together with IL-6 and soluble IL-6 receptor. IL6RIL6 chimera prolonged in vitro maintenance of SRC for up to 14 days. Stimulation of CD34+CD38−/low enriched cells with IL6RIL6 chimera maintained the early CD34+CD38−/lowcell subpopulation, which could be detected in vitro for up to 14 days. Moreover, IL6RIL6 chimera preferentially stimulated the growth of early CD34+38−/low cells, resulting in significantly higher levels of progenitors compared with more mature CD34+38+ cells. Taken together, these findings demonstrate the importance of IL6RIL6 chimera in stimulating the proliferation of early CD34+· CD38−gp130+IL-6R−cells in vitro and extended maintenance of progenitors and SRC.
T-Cell Receptor-Independent Activation of Clonal Th2 Cells Associated With Chronic Hypereosinophilia
Authors: Roufosse Florence , Schandené Liliane , Sibille Catherine , Kennes Bernard , Efira André , Cogan Elie , Goldman Michel
Published: October 2019

We recently observed a clonal expansion of CD3−CD4+ T cells secreting Th2-type cytokines in patients presenting chronic hypereosinophilia. As clonal T cells isolated from such patients did not spontaneously secrete cytokines in vitro, we reasoned that costimulatory signals delivered by antigen-presenting cells might be required to induce their full activation. To address this question, we investigated in two such patients the responses of CD3−CD4+ T cells to dendritic cells (DC). DC elicited proliferation and production of interleukin-5 (IL-5) and IL-13 by clonal cells from patient 1 and upregulated their expression of CD25 (IL-2R-). These effects were abolished when blocking monoclonal antibodies (MoAbs) against IL-2R- and IL-2 were added to cocultures, indicating critical involvement of an autocrine IL-2/IL-2R pathway. Cells from patient 2 were stimulated by DC to produce Th2 cytokines only when rIL-2 or rIL-15 was added to cocultures. In both patients, addition of inhibitory MoAbs against B7-1/B7-2 or CD2 to cocultures resulted in dramatic reduction of cytokine production and inhibited CD25 upregulation. Thus, TCR/CD3-independent activation of clonal Th2 cells by DC is an IL-2–dependent process, which requires signaling through CD2 and CD28.
Homing of Human Cells in the Fetal Sheep Model: Modulation by Antibodies Activating or Inhibiting Very Late Activation Antigen-4–Dependent Function
Authors: Zanjani Esmail D. , Flake Alan W. , Almeida-Porada Graça , Tran Nam , Papayannopoulou Thalia
Published: October 2019

The mechanisms by which intravenously (IV)-administered hematopoietic cells home to the bone marrow (BM) are poorly defined. Although insightful information has been obtained in mice, our knowledge about homing of human cells is very limited. In the present study, we investigated the importance of very late activation antigen (VLA)-4 in the early phases of lodgment of human CD34+progenitors into the sheep hematopoietic compartment after in utero transplantation. We have found that preincubation of donor cells with anti–VLA-4 blocking antibodies resulted in a profound reduction of human cell lodgment in the fetal BM at 24 and 48 hours after transplantation, with a corresponding increase of human cells in the peripheral circulation. Furthermore, IV infusion of the anti–VLA-4 antibody at later times (posttransplantation days 21 to 24) resulted in redistribution or mobilization of human progenitors from the BM to the peripheral blood. In an attempt to positively modulate homing, we also pretreated human donor cells with an activating antibody to β1 integrins. This treatment resulted in increased lodgment of donor cells in the fetal liver, presumably for hemodynamic reasons, at the expense of the BM. Given previous involvement of the VLA-4/vascular cell adhesion molecule (VCAM)-1 adhesion pathway in homing and mobilization in the murine system, our present data suggest that cross-reacting ligands (likely VCAM-1) for human VLA-4 exist in sheep BM, thereby implicating conservation of molecular mechanisms of homing and mobilization across disparate species barriers. Thus, information from xenogeneic models of human hematopoiesis and specifically, the human/sheep model of in utero transplantation, may provide valuable insights into human hematopoietic transplantation biology.
MUC1 dysregulation as the consequence of a t(1;14)(q21;q32) translocation in an extranodal lymphoma
Authors: Gilles Frédéric , Goy André , Remache Yvonne , Shue Peter , Zelenetz Andrew D.
Published: October 2019

Cytogenetic abnormalities at chromosome 1q21 are among the most common lesions in diffuse large-cell lymphoma and have been associated with a poor prognosis. A novel cell line, SKI-DLCL-1, was established from ascitic fluid that carries a t(1;14)(q21;q32) chromosomal translocation. Using pulsed-field gel electrophoresis, the breakpoint on the IgH locus mapped to a gamma locus between C1 and C2. A cosmid library was prepared from SKI-DLCL-1, and Cγ-positive clones spanning the breakpoint were identified by screening with fluorescence in situ hybridization. The breakpoint occurs 860 bp downstream of the 3′ UTR of the MUC1 gene. The break appears to be a staggered double-strand break consistent with an error in immunoglobulin class switching. The MUC1 gene is highly transcribed and translated, and the protein is highly glycosylated. It is postulated that MUC1 expression is brought under the control of the 3′E enhancer. MUC1 lies in a region of chromosome 1 characterized by an unusually high density of genes, with 7 known genes in a region of approximately 85 kb. To determine whether there was a pleiotropic effect of the expression of genes in the region as a consequence of the translocation, the expression of 6 additional genes was assessed. None of the other genes in this region (CLK2, propin, COTE1, GBA, metaxin, and thrombospondin 3) are overexpressed in SKI-DLCL-1. Thus, the translocation t(1;14)(q21;q32) seen in both the primary tumor and the derived cell line results in the marked overexpression of MUC1 without affecting the expression of other genes in the region.
Oral isobutyramide reduces transfusion requirements in some patients with homozygous β-thalassemia
Authors: Reich Susanne , Bührer Christoph , Henze Günter , Ohlendorf Dieter , Mesche Michael , Sinha Pranav , Kage Andreas , Müller Christian , Vetter Barbara , Kulozik Andreas E.
Published: October 2019

The butyrate derivative isobutyramide (IBT) increases fetal hemoglobin (HbF) in patients with β-hemoglobinopathies, but little is known about its usefulness for prolonged therapeutic use. We treated 8 patients with transfusion-dependent β-thalassemia with 350 mg/kg of body weight per day of oral IBT for 126 to 384 days. During the trial period, the hemoglobin level was maintained between 85 g/L (range 82-87 g/L) (pretransfusion) and 115 g/L (range 110-119 g/L) (post-transfusion) (median, interquartile range), corresponding to 4-week transfusion intervals in all patients during the pretreatment phase. Adverse effects (bitter taste, epigastric discomfort) did not cause discontinuation of IBT. HbF increased in all patients from 3.1% (range 1.9%-4.8%) to 6.0% (range 3.3%-8.7) (P = .0017), while free Hb dropped from 0.48 g/L (range 0.39-0.81 g/L) to 0.19 g/L (range 0.16-0.24 g/L) (P < .0001). Transfusion intervals were consistently extended to 8 or 9 weeks in 1 patient, resulting in a decrease of daily iron load from 455 μg/kg per day (range 451-459 μg/kg per day) before therapy to 211μg/kg per day (range 203-286 μg/kg per day) during the 12-month treatment period. Prolongation of transfusion intervals achieved by IBT was less consistent in another patient, whose parenteral iron load nevertheless decreased from 683 μg/kg per day (range 618-748 μg/kg per day) to 542 μg/kg per day (340-596 μg/kg per day). In the other 6 patients, no prolongation of transfusion intervals was achieved. Response to treatment was associated with high pretreatment HbF (> 4.5%), high parental HbF, and increased erythropoietin levels (> 150 IU/L). We conclude that IBT prolongs transfusion intervals and reduces parenteral iron burden in some patients with transfusion-dependent β-thalassemia.
Differential surrogate light chain expression governs B-cell differentiation
Authors: Wang Yui-Hsi , Stephan Robert P. , Scheffold Alexander , Kunkel Désirée , Karasuyama Hajime , Radbruch Andreas , Cooper Max D.
Published: October 2019

Surrogate light chain expression during B lineage differentiation was examined by using indicator fluorochrome-filled liposomes in an enhanced immunofluorescence assay. Pro-B cells bearing surrogate light chain components were found in mice, but not in humans. A limited subpopulation of relatively large pre-B cells in both species expressed pre-B cell receptors. These cells had reduced expression of the recombinase activating genes, RAG-1 and RAG-2. Their receptor-negative pre-B cell progeny were relatively small, expressed RAG-1 and RAG-2, and exhibited selective down-regulation of VpreB and λ5expression. Comparative analysis of the 2 pre-B cell subpopulations indicated that loss of the pre-B cell receptors from surrogate light chain gene silencing was linked with exit from the cell cycle and light chain gene rearrangement to achieve B-cell differentiation.
Long-term control of recurrent or refractory viral infections after allogeneic HSCT with third-party virus-specific T cells
Authors: Withers Barbara , Blyth Emily , Clancy Leighton E. , Yong Agnes , Fraser Chris , Burgess Jane , Simms Renee , Brown Rebecca , Kliman David , Dubosq Ming-Celine , Bishop David , Sutrave Gaurav , Ma Chun Kei Kris , Shaw Peter J. , Micklethwaite Kenneth P. , Gottlieb David J.
Published: October 2019

Key Points

Partially HLA-matched third-party CMV-specific T cells provide long-term viral control in HSCT patients with resistant CMV infection. Viral control occurs in the setting of recovery of CD8+ terminally differentiated effector T cells.
A novel method to generate T-cell receptor–deficient chimeric antigen receptor T cells
Authors: Kamiya Takahiro , Wong Desmond , Png Yi Tian , Campana Dario
Published: October 2019

Key Points

Newly designed PEBLs prevent surface expression of T-cell receptor in T cells without affecting their function. Combined with chimeric antigen receptors, PEBLs can rapidly generate powerful antileukemic T cells without alloreactivity.
Rates and predictors of switching to tenofovir alafenamide-containing ART in a nationwide cohort
Authors:   the Swiss HIV Cohort Study, Surial Bernard , Cavassini Matthias , Calmy Alexandra , Fehr Jan , Stöckle Marcel , Bernasconi Enos , Roth Bianca , Fux Christoph A. , Kovari Helen , Furrer Hansjakob , Rauch Andri , Wandeler Gilles
Published: October 2019

Background

Tenofovir alafenamide (TAF)-containing combinations were introduced in Switzerland after October 2016 and are recommended over tenofovir disoproxil fumarate (TDF) in patients with osteoporosis or impaired renal function.

Methods

We included all participants of the Swiss HIV Cohort Study on TDF-containing antiretroviral therapy with follow-up visits after January 2016. We determined the proportion of switches from TDF to TAF overall, and among patients with risk factors for TDF toxicity, including osteoporosis, impaired renal function or marked proteinuria. We used multivariable logistic regression to explore predictors of switching from TDF to TAF.

Results

We included 5′012 patients, of whom 652 (13.0%) had risk factors for TDF toxicity. A switch from TDF to TAF was undertaken in 2′796 (55.8%) individuals overall, and in 465 (71.3%) with risk factors. Predictors of switching to TAF were male sex (adjusted odds ratio 1.27, 95% confidence interval 1.07–1.50), age > 50 years (1.43, 1.23–1.66) and the presence of risk factors for TDF toxicity (2.21, 1.77–2.75). In contrast, patients with a non-nucleoside reverse transcriptase inhibitor (NNRTI)-based single-pill regimen (0.11, 0.09–0.13), those treated in non-tertiary care centers (0.56, 0.46–0.70), as well as those with CD4 cell counts below 500/μL (0.77, 0.66–0.90) and with chronic hepatitis C infection (0.66, 0.54–0.80) were most likely to stay on TDF.

Conclusions

Over 50% of patients on TDF-containing therapy, including the majority of patients at risk for TDF toxicity, were switched to TAF within two years of its introduction in Switzerland. Individuals on NNRTI-based single-pill regimens were most likely to remain on TDF.
Dorsal subscapularis approach for the surgical drainage of subscapularis intramuscular abscess: a case report
Authors: Furuhata Ryogo , Inoue Doji , Kiyota Yasuhiro , Morioka Hideo , Arino Hiroshi
Published: October 2019

Background

Abscess formation in the subscapularis muscle is a rare clinical condition. Few reports are available regarding the treatment methods and surgical approaches for subscapularis intramuscular abscesses. Here, we describe a case of subscapularis intramuscular abscess that was treated successfully via surgical drainage using a new approach, the “dorsal subscapularis approach”.

Case presentation

A 67-year-old woman presented to our hospital with complaints of fever and disturbance of consciousness. Two days prior to visiting our hospital, right shoulder pain and limited range of motion in the shoulder were noted. Cerebrospinal fluid examination and contrast-enhanced computed tomography (CT) imaging on admission revealed a right subscapularis intramuscular abscess with concomitant bacterial meningitis. The patient’s clinical symptoms improved after antibiotic administration for 3 weeks, but the right shoulder pain persisted. Contrast-enhanced CT imaging performed after antibiotic administration revealed an abscess in the right shoulder joint space, in addition to a capsule of the abscess in the right subscapularis muscle. We performed open surgical drainage for the abscess, which had spread from the subscapularis muscle to the glenohumeral joint. Using the deltoid-pectoral approach, we detected exudate and infected granulation tissue in the joint cavity. Furthermore, we separated the dorsal side of the subscapularis muscle from the scapula using a raspatory and detected infected granulation tissue in the subscapularis muscle belly. We performed curettage and washed as much as possible. After surgery, antibiotic administration continued for 2 weeks. The patient’s right shoulder pain subsided and CT performed 2 months after surgery revealed no recurrence of infection.

Conclusions

The present case indicated that a subscapularis intramuscular abscess could lead to severe concomitant infections of other organs via the hematogenous route. Thus, early detection and treatment are necessary. Moreover, in this case, surgical drainage using a dorsal subscapularis approach was beneficial to treating the abscess, which had spread from the subscapularis muscle to the glenohumeral joint.
Volumetric characteristics of idiopathic pulmonary fibrosis lungs: computational analyses of high-resolution computed tomography images of lung lobes
Authors: Sul Bora , Flors Lucia , Cassani Joanne , Morris Michael J. , Reifman Jaques , Altes Talissa , Wallqvist Anders
Published: October 2019

Background

Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease involving progressive degeneration of lung capacity. Current diagnosis of IPF heavily relies on visual evaluation of fibrotic features in high-resolution computed tomography (HRCT) images of the lungs. Although the characteristics of this disease have been studied at the molecular and cellular levels, little is known about the mechanical characteristics of IPF lungs inferred from HRCT images. To this end, we performed a pilot study to investigate the radiographic and volumetric characteristics of lungs in subjects with IPF.

Methods

We collected HRCT images of healthy (N = 13) and IPF (N = 9) lungs acquired at breath-holds after full inspiration (expanded state) and full expiration (contracted state). We performed statistical analyses on Hounsfield unit (HU) histograms, lobar volumes (V: lobe volume normalized by the lung volume), and lobar flows (Q: the difference in lobe volume divided by the difference in lung volume between the expanded and contracted states).

Results

Parameters characterizing the HU histograms (i.e., mean, median, skewness, and kurtosis) significantly differed between healthy and IPF subjects, for all lobes in both expanded and contracted states. The distribution ofVacross lobes differed significantly between the groups in both states. The distribution ofQalso differed significantly between the groups:Qvalues of the lower lobes for the IPF group were 33% (right) and 22% (left) smaller than those for the healthy group, consistent with the observation that radiographic scores were highest in the lower lung section in IPF. Notably, the root-mean-squared difference (RMSD) ofQ, a measure of distance from the mean value of the healthy group, clearly distinguished the IPF subjects (RMSD ofQ > 1.59) from the healthy group (RMSD ofQ < 0.67).

Conclusion

This study shows that lung volume and flow distribution change heterogeneously across the lung lobes of IPF subjects, with reduced capacity in the lower lobes. These volumetric changes may improve our understanding of the pathophysiology in IPF lungs.
Construction of a novel MK-4 biosynthetic pathway in Pichia pastoris through heterologous expression of HsUBIAD1
Authors: Sun Xiaowen , Liu Hui , Wang Peng , wang Li , Ni Wenfeng , Yang Qiang , Wang Han , Tang Hengfang , Zhao Genhai , Zheng Zhiming
Published: October 2019

Background

With a variety of physiological and pharmacological functions, menaquinone is an essential prenylated product that can be endogenously converted from phylloquinone (VK1) or menadione (VK3) via the expression ofHomo sapiensUBIAD1 (HsUBIAD1). The methylotrophic yeast,Pichia pastoris, is an attractive expression system that has been successfully applied to the efficient expression of heterologous proteins. However, the menaquinone biosynthetic pathway has not been discovered inP. pastoris.

Results

Firstly, we constructed a novel synthetic pathway inP. pastorisfor the production of menaquinone-4 (MK-4) via heterologous expression ofHsUBIAD1. Then, the glyceraldehyde-3-phosphate dehydrogenase constitutive promoter (PGAP) appeared to be mostsuitable for the expression ofHsUBIAD1 for various reasons. By optimizing the expression conditions ofHsUBIAD1, its yield increased by 4.37 times after incubation at pH 7.0 and 24 °C for 36 h, when compared with that under the initial conditions. We foundHsUBIAD1 expressed in recombinant GGU-23 has the ability to catalyze the biosynthesis of MK-4 when using VK1and VK3as the isopentenyl acceptor. In addition, we constructed a ribosomal DNA (rDNA)-mediated multi-copy expression vector for the fusion expression ofSaGGPPS andPpIDI, and the recombinant GGU-GrIG afforded higher MK-4 production, so that it was selected as the high-yield strain. Finally, the yield of MK-4 was maximized at 0.24 mg/g DCW by improving the GGPP supply when VK3was the isopentenyl acceptor.

Conclusions

In this study, we constructed a novel synthetic pathway inP. pastorisfor the biosynthesis of the high value-added prenylated product MK-4 through heterologous expression ofHsUBIAD1 and strengthened accumulation of GGPP. This approach could be further developed and accomplished for the biosynthesis of other prenylated products, which has great significance for theoretical research and industrial application.
Prognostic value of YKL-40 in solid tumors: a meta-analysis of 41 cohort studies
Authors: Bian Bingxian , Li Li , Yang Junyao , Liu Yi , Xie Guohua , Zheng Yingxia , Zeng Liang , Zeng Junxiang , Shen Lisong
Published: October 2019

Background

Serum/plasma YKL-40 can be a useful index that is associated with tumor development. However, the prognostic value of serum/plasma YKL-40 in patients with solid tumors is still unclear. We aimed to utilize the existing literature to investigate the prognostic value of serum/plasma YKL-40 in solid tumors.

Methods

An extensive literature search for relevant studies was conducted with the Embase, Medline and Web of Science databases. The effect on survival was measured with the hazard ratio (HR). Then, pooled HRs and 95% confidence intervals (CIs) were calculated using the random and fixed-effects models according to the heterogeneity of the included studies.

Results

This meta-analysis was based on 41 publications and comprised a total of 7762 patients with solid tumors. The pooled HR showed that elevated serum/plasma YKL-40 was significantly associated with poor OS (HR, 1.44; 95% CI 1.33–1.56). We also found that elevated serum/plasma YKL-40 had significant prognostic effects on OS in various cancer subgroups such as gastrointestinal tumors (HR, 1.37; 95% CI 1.18–1.58), ovarian cancer (HR, 2.27; 95% CI 1.69–3.06), melanoma (HR, 1.77; 95% CI 1.18–2.67), lung cancer (HR, 1.73; 95% CI 1.35–2.23), urologic neoplasms (HR, 1.61; 95% CI 1.08–2.40) and glioblastoma (HR, 1.23; 95% CI 1.07–1.42); in contrast, the prognostic effect of serum/plasma YKL-40 was not statistically significant in breast cancer (HR, 1.07; 95% CI 0.98–1.17).

Conclusions

The available evidence supports the hypothesis that elevated serum/plasma YKL-40 is associated with poor survival in patients with solid tumors and that serum/plasma YKL-40 may serve as a novel prognostic biomarker.
Identification of Cancer–associated metabolic vulnerabilities by modeling multi-objective optimality in metabolism
Authors: Dai Ziwei , Yang Shiyu , Xu Liyan , Hu Hongrong , Liao Kun , Wang Jianghuang , Wang Qian , Gao Shuaishi , Li Bo , Lai Luhua
Published: October 2019

Background

Cancer cells undergo global reprogramming of cellular metabolism to satisfy demands of energy and biomass during proliferation and metastasis. Computational modeling of genome-scale metabolic models is an effective approach for designing new therapeutics targeting dysregulated cancer metabolism by identifying metabolic enzymes crucial for satisfying metabolic goals of cancer cells, but nearly all previous studies neglect the existence of metabolic demands other than biomass synthesis and trade-offs between these contradicting metabolic demands. It is thus necessary to develop computational models covering multiple metabolic objectives to study cancer metabolism and identify novel metabolic targets.

Methods

We developed a multi-objective optimization model for cancer cell metabolism at genome-scale and an integrated, data-driven workflow for analyzing the Pareto optimality of this model in achieving multiple metabolic goals and identifying metabolic enzymes crucial for maintaining cancer-associated metabolic phenotypes. Using this workflow, we constructed cell line-specific models for a panel of cancer cell lines and identified lists of metabolic targets promoting or suppressing cancer cell proliferation or the Warburg Effect. The targets were then validated using knockdown and over-expression experiments in cultured cancer cell lines.

Results

We found that the multi-objective optimization model correctly predicted phenotypes including cell growth rates, essentiality of metabolic genes and cell line specific sensitivities to metabolic perturbations. To our surprise, metabolic enzymes promoting proliferation substantially overlapped with those suppressing the Warburg Effect, suggesting that simply targeting the overlapping enzymes may lead to complicated outcomes. We also identified lists of metabolic enzymes important for maintaining rapid proliferation or high Warburg Effect while having little effect on the other. The importance of these enzymes in cancer metabolism predicted by the model was validated by their association with cancer patient survival and knockdown and overexpression experiments in a variety of cancer cell lines.

Conclusions

These results confirm this multi-objective optimization model as a novel and effective approach for studying trade-off between metabolic demands of cancer cells and identifying cancer-associated metabolic vulnerabilities, and suggest novel metabolic targets for cancer treatment.

Graphical abstract
MeCP2-E1 isoform is a dynamically expressed, weakly DNA-bound protein with different protein and DNA interactions compared to MeCP2-E2
Authors: Martínez de Paz Alexia , Khajavi Leila , Martin Hélène , Claveria-Gimeno Rafael , Tom Dieck Susanne , Cheema Manjinder S. , Sanchez-Mut Jose V. , Moksa Malgorzata M. , Carles Annaick , Brodie Nick I. , Sheikh Taimoor I. , Freeman Melissa E. , Petrotchenko Evgeniy V. , Borchers Christoph H. , Schuman Erin M. , Zytnicki Matthias , Velazquez-Campoy Adrian , Abian Olga , Hirst Martin , Esteller Manel , Vincent John B. , Malnou Cécile E. , Ausió Juan
Published: October 2019

Background

MeCP2—a chromatin-binding protein associated with Rett syndrome—has two main isoforms, MeCP2-E1 and MeCP2-E2, differing in a few N-terminal amino acid residues. Previous studies have shown brain region-specific expression of these isoforms which, in addition to their different cellular localization and differential expression during brain development, suggest that they may also have non-overlapping molecular mechanisms. However, differential functions of MeCP2-E1 and E2 remain largely unexplored.

Results

Here, we show that the N-terminal domains (NTD) of MeCP2-E1 and E2 modulate the ability of the methyl-binding domain (MBD) to interact with DNA as well as influencing the turn-over rates, binding dynamics, response to neuronal depolarization, and circadian oscillations of the two isoforms. Our proteomics data indicate that both isoforms exhibit unique interacting protein partners. Moreover, genome-wide analysis using ChIP-seq provide evidence for a shared as well as a specific regulation of different sets of genes.

Conclusions

Our study supports the idea that Rett syndrome might arise from simultaneous impairment of cellular processes involving non-overlapping functions of MECP2 isoforms. For instance, MeCP2-E1 mutations might impact stimuli-dependent chromatin regulation, while MeCP2-E2 mutations could result in aberrant ribosomal expression. Overall, our findings provide insight into the functional complexity of MeCP2 by dissecting differential aspects of its two isoforms.
Role of neuromedin B and its receptor in the innate immune responses against influenza A virus infection in vitro and in vivo
Authors: Yang Guihong , Huang Huipeng , Tang Mengyao , Cai Zifeng , Huang Cuiqin , Qi Baomin , Chen Ji-Long
Published: October 2019

The peptide neuromedin B (NMB) and its receptor (NMBR) represent a system (NMB/NMBR) of neuromodulation. Here, it was demonstrated that the expression of NMBR in cells or murine lung tissues was clearly upregulated in response to H1N1/PR8 influenza A virus infection. Furthermore, the in vitro and in vivo activities of NMB/NMBR during PR8 infection were investigated. It was observed that A549 cells lacking endogenous NMBR were more susceptible to virus infection than control cells, as evidenced by the increased virus production in the cells. Interestingly, a significant decrease in IFN-α and increased IL-6 expression were observed in these cells. The role of this system in innate immunity against PR8 infection was probed by treating mice with NMB. The NMB-treated mice were less susceptible to virus challenge, as evidenced by increased survival, increased body weight, and decreased viral NP expression compared with the control animals. Additionally, the results showed that exogenous NMB not only enhanced IFN-α expression but also appeared to inhibit the expression of NP and IL-6 in PR8-infected cells and animals. As expected, opposing effects were observed in the NMBR antagonist-treated cells and mice, which further confirmed the effects of NMB. Together, these data suggest that NMB/NMBR may be an important component of the host defence against influenza A virus infection. Thus, these proteins may serve as promising candidates for the development of novel antiviral drugs.
The effect of vitamin D supplementation on the glycemic control of pre-diabetic Qatari patients in a randomized control trial
Authors: Al Thani Mohammed , Sadoun Eman , Sofroniou Angeliki , Jayyousi Amin , Baagar Khaled Ahmed Mohamed , Al Hammaq Abdulla , Vinodson Benjamin , Akram Hammad , Bhatti Zaid Shakoor , Nasser Heba Samir , Leventakou Vasiliki
Published: October 2019

Background

Vitamin D deficiency is associated with indicators of pre-diabetes including, insulin resistance, β-cell dysfunction and elevated plasma glucose with controversial findings from current trials. This study aims to investigate the long-term effect of vitamin D on glucose metabolism and insulin sensitivity in pre-diabetic and highly vitamin-deficient subjects.

Methods

One hundred thirty-two participants were randomized to 30,000 IU vitamin D weekly for 6 months. Participants underwent oral glucose tolerance test (OGTT) at 3-month intervals to determine the change in plasma glucose concentration at 2 h after 75 g OGTT (2hPCG). Secondary measurements included glycated hemoglobin, fasting plasma glucose and insulin, post-prandial insulin, indices of insulin sensitivity (HOMA-IR, Matsuda Index), β-cell function (HOMA-β, glucose and insulin area under the curve (AUC), disposition and insulinogenic indices), and lipid profile.

Results

A total of 57 (vitamin D) and 75 (placebo) subjects completed the study. Mean baseline serum 25(OH) D levels were 17.0 ng/ml and 14.9 ng/ml for placebo and vitamin D group, respectively. No significant differences were observed for 2hPC glucose or insulin sensitivity indices between groups. HOMA-β significantly decreased in the vitamin D group, while area under curve for glucose and insulin showed a significant reduction in β-cell function in both groups. Additionally, HOMA-β was found to be significantly different between control and treatment group and significance persisted after adjusting for confounding factors.

Conclusion

Vitamin D supplementation in a pre-diabetic and severely vitamin-deficient population had no effect on glucose tolerance or insulin sensitivity. The observed reduction in β-cell function in both placebo and vitamin D groups could be attributed to factors other than supplementation.

Trial registration

NCT02098980, 28/03/2014 (www.clinicaltrials.gov).
Adapting and implementing Caring Contacts in a Department of Veterans Affairs emergency department: a pilot study protocol
Authors: Landes Sara J. , Kirchner JoAnn E. , Areno John P. , Reger Mark A. , Abraham Traci H. , Pitcock Jeffery A. , Bollinger Mary J. , Comtois Katherine Anne
Published: October 2019

Background

Suicide among veterans is a problem nationally, and suicide prevention remains a high priority for the Department of Veterans Affairs (VA). Focusing suicide prevention initiatives in the emergency department setting provides reach to veterans who may not be seen in mental health and targets a critical risk period, transitions in care following discharge. Caring Contacts is a simple and efficacious suicide prevention approach that could be used to target this risk period. The purpose of this study is to (1) adapt Caring Contacts for use in a VA emergency department, (2) conduct a pilot program at a single VA emergency department, and (3) create an implementation toolkit to facilitate spread of Caring Contacts to other VA facilities.

Methods

This project includes planning activities and a pilot at a VA emergency department. Planning activities will include determining available data sources, determining logistics for identifying and sending Caring Contacts, and creating an implementation toolkit. We will conduct qualitative interviews with emergency department staff and other key stakeholders to gather data on what is needed to adapt and implement Caring Contacts in a VA emergency department setting and possible barriers to and facilitators of implementation. An advisory board of key stakeholders in the facility will be created. Qualitative findings from interviews will be presented to the advisory board for discussion, and the board will use these data to inform decision making regarding implementation of the pilot. Once the pilot is underway, the advisory board will convene again to discuss ongoing progress and determine if any changes are needed to the implementation of the Caring Contacts intervention.

Discussion

Findings from the current project will inform future scale-up and spread of this innovation to other VA medical center emergency departments across the network and other networks. The current pilot will adapt Caring Contacts, create an implementation toolkit and implementation guide, evaluate the feasibility of gathering outcome measures, and provide information about what is needed to implement this evidence-based suicide prevention intervention in a VA emergency department.
A nomogram predicting the recurrence of hepatocellular carcinoma in patients after laparoscopic hepatectomy
Authors: Pan Yang‐Xun , Chen Jian‐Cong , Fang Ai‐Ping , Wang Xiao‐Hui , Chen Jin‐Bin , Wang Jun‐Cheng , He Wei , Fu Yi‐Zhen , Xu Li , Chen Min‐Shan , Zhang Yao‐Jun , Li Qi‐Jiong , Zhou Zhong‐Guo
Published: October 2019

Background

Patients with hepatocellular carcinoma (HCC) undergoing surgical resection still have a high 5‐year recurrence rate (~ 60%). With the development of laparoscopic hepatectomy (LH), few studies have compared the efficacy between LH and traditional surgical approach on HCC. The objective of this study was to establish a nomogram to evaluate the risk of recurrence in HCC patients who underwent LH.

Methods

The clinical data of 432 patients, pathologically diagnosed with HCC, underwent LH as initial treatment and had surgical margin > 1 cm were collected. The significance of their clinicopathological features to recurrence‐free survival (RFS) was assessed, based on which a nomogram was constructed using a training cohort (n = 324) and was internally validated using a temporal validation cohort (n = 108).

Results

Hepatitis B surface antigen (hazard ratio [HR], 1.838; P= 0.044), tumor number (HR, 1.774; P= 0.003), tumor thrombus (HR, 2.356; P= 0.003), cancer cell differentiation (HR, 0.745; P= 0.080), and microvascular tumor invasion (HR, 1.673; P =0.007) were found to be independent risk factors for RFS in the training cohort, and were used for constructing the nomogram. The C‐index for RFS prediction in the training cohort using the nomogram was 0.786, which was higher than that of the 8th edition of the American Joint Committee on Cancer TNM classification (C‐index, 0.698) and the Barcelona Clinic Liver Cancer staging system (C‐index, 0.632). A high consistency between the nomogram prediction and actual observation was also demonstrated by a calibration curve. An improved predictive benefit in RFS and higher threshold probability of the nomogram were determined by receiver operating characteristic curve analysis, which was also confirmed in the validation cohort compared to other systems.

Conclusions

We constructed and validated a nomogram able to quantify the risk of recurrence after initial LH for HCC patients, which can be clinically implemented in assisting the planification of individual postoperative surveillance protocols.
The Brazilian Society of Rheumatology recommendations on investigation and diagnosis of systemic autoimmune myopathies
Authors: de Souza Fernando Henrique Carlos , de Araújo Daniel Brito , Vilela Verônica Silva , Simões Ricardo Santos , Bernardo Wanderley Marques , Frank Thais Amanda , da Cunha Bernardo Matos , Shinjo Samuel Katsuyuki
Published: October 2019

Background

This research is recommended by the Myopathy Committee of the Brazilian Society of Rheumatology for the investigation and diagnosis of systemic autoimmune myopathies.

Body

A systematic literature review was performed in the Embase, Medline (PubMed) and Cochrane databases, including studies published until October 2018. PRISMA was used for the review, and the articles were evaluated, based on the Oxford levels of evidence. Ten recommendations were developed addressing different aspects of systemic autoimmune myopathy investigation and diagnosis.

Conclusions

The European League Against Rheumatism/ American College of Rheumatology (EULAR/ACR) classification stands out for the diagnosis of systemic autoimmune myopathies. Muscular biopsy is essential, aided by muscular magnetic resonance images and electroneuromyography in complementary research. Analysis of the factors related to prognosis with the evaluation of extramuscular manifestations, and comorbidities and intense investigation regarding differential diagnoses are mandatory.
Management of clozapine-induced sialorrhoea
Authors: Gupta Sumeet , Khastgir Udayan , Croft Matthew , Roshny Sefat
Published: October 2019

Sialorrhoea (hypersalivation) is a common adverse effect of clozapine. If severe, it can affect patients' quality of life and adherence to the treatment. Clinicians therefore need to proactively manage this effect. At present, no drugs are licensed to manage clozapine-induced sialorrhoea, although there are many off-label treatment options, with variable effectiveness. Anticholinergic medications are commonly prescribed for it, but they have limited effect and can worsen constipation. This article gives a brief overview of other practical and pharmacological management options.
Inflammatory Stimuli Trigger Increased Androgen Production and Shifts in Gene Expression in Theca-Interstitial Cells
Authors: Fox Chelsea W , Zhang Lingzhi , Sohni Abhishek , Doblado Manuel , Wilkinson Miles F , Chang R Jeffrey , Duleba Antoni J
Published: October 2019

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by theca cell hyperplasia and excessive androgen production. An increasing body of evidence has pointed to a close association between PCOS and low-grade chronic systemic inflammation. However, the mechanistic basis for this linkage is unknown. Therefore, we evaluated the effects of the inflammatory agents lipopolysaccharide (LPS) and IL-1β on rat theca-interstitial cells (TICs). We found that incubation with either LPS or IL-1β elicited a dose-dependent increase in both TIC viability and androgen production. Using RNA sequencing analysis, we found that both of these inflammatory agents also triggered profound and widespread shifts in gene expression. Using a stringent statistical cutoff, LPS and IL-1β elicited differential expression of 5201 and 5953 genes, respectively. Among the genes upregulated by both LPS and IL-1β were key regulatory genes involved in the cholesterol and androgen biosynthesis pathways, including Cyp17a1, Cyp11a1, Hsd3b, and Hmgcr. This provides a molecular explanation for the mechanism of action of inflammatory agents leading to increased androgen production. Gene ontology and pathway analysis revealed that both LPS and IL-1β regulated genes highly enriched for many common functions, including the immune response and apoptosis. However, a large number of genes (n = 2222) were also uniquely regulated by LPS and IL-1β, indicating that these inflammatory mediators have substantial differences in their mechanism of action. Together, these findings highlight the potential molecular mechanisms through which chronic low-grade inflammation contributes to the pathogenesis of androgen excess in PCOS.
Cell cycle-independent furrowing triggered by phosphomimetic mutations of the INCENP STD motif requires Plk1
Authors: Papini Diana , Fant Xavier , Ogawa Hiromi , Desban Nathalie , Samejima Kumiko , Feizbakhsh Omid , Askin Bilge , Ly Tony , Earnshaw William C. , Ruchaud Sandrine
Published: October 2019

Timely and precise control of Aurora B kinase, the chromosomal passenger complex (CPC) catalytic subunit, is essential for accurate chromosome segregation and cytokinesis. Post-translational modifications of CPC subunits are directly involved in controlling Aurora B activity. Here, we identified a highly conserved acidic STD-rich motif of INCENP that is phosphorylated during mitosis in vivo and by Plk1 in vitro and is involved in controlling Aurora B activity. By using an INCENP conditional-knockout cell line, we show that impairing the phosphorylation status of this region disrupts chromosome congression and induces cytokinesis failure. In contrast, mimicking constitutive phosphorylation not only rescues cytokinesis but also induces ectopic furrows and contractile ring formation in a Plk1- and ROCK1-dependent manner independent of cell cycle and microtubule status. Our experiments identify the phospho-regulation of the INCENP STD motif as a novel mechanism that is key for chromosome alignment and cytokinesis.

This article has an associated First Person interview with the first author of the paper.
The Relationship Between Tumor Budding, Tumor Microenvironment, and Survival in Patients with Primary Operable Colorectal Cancer
Authors: van Wyk Hester C. , Roseweir Antonia , Alexander Peter , Park James H. , Horgan Paul G. , McMillan Donald C. , Edwards Joanne
Published: October 2019

Background

Tumor budding is an independent prognostic factor in colorectal cancer (CRC) and has recently been well-defined by the International Tumour Budding Consensus Conference (ITBCC).

Objective

The aim of the present study was to use the ITBCC budding evaluation method to examine the relationship between tumor budding, tumor factors, tumor microenvironment, and survival in patients with primary operable CRC.

Methods

Hematoxylin and eosin-stained slides of 952 CRC patients diagnosed between 1997 and 2007 were evaluated for tumor budding according to the ITBCC criteria. The tumor microenvironment was evaluated using tumor stroma percentage (TSP) and Klintrup–Makinen (KM) grade to assess the tumor inflammatory cell infiltrate.

Results

High budding (n = 268, 28%) was significantly associated with TNM stage (p < 0.001), competent mismatch repair (MMR;p < 0.05), venous invasion (p < 0.001), weak KM grade (p < 0.001), high TSP (p < 0.001), and reduced cancer-specific survival (CSS) (hazard ratio 8.68, 95% confidence interval 6.30–11.97;p < 0.001). Tumor budding effectively stratifies CSS stage T1 through to T4 (allp < 0.05) independent of associated factors.

Conclusions

Tumor budding effectively stratifies patients’ survival in primary operable CRC independent of other phenotypic features. In particular, the combination of T stage and budding should form the basis of a new staging system for primary operable CRC.
Exploring the impact of glutathione S-transferase (GST)-based metabolic resistance to insecticide on vector competence of Anopheles funestus for Plasmodium falciparum
Authors: Ndo Cyrille , Kopya Edmond , Irving Helen , Wondji Charles
Published: October 2019

Background:Malaria control heavily relies on insecticide-based interventions against mosquito vectors. However, the increasing spread of insecticide resistance is a major threat. The extent to which such resistance, notably metabolic resistance, influences the development of thePlasmodiumparasite and its impact on overall malaria transmission remains poorly characterized. Here, we investigated whether glutathione S-transferase-based resistance could influencePlasmodium falciparumdevelopment inAnopheles funestus.

Methods:Anopheles funestusfemales were infected withP. falciparumgametocytes and midguts were dissected at day 7 post infection for detection/quantification of oocysts. Infection parameters were compared between individuals with different L119F-GSTe2 genotypes, and the polymorphism of the GSTe2 gene was analyzed in infected and uninfected mosquito groups.

Results:Overall, 403An. funestus mosquitoes were dissected and genotyped. The frequency of the L119F-GSTe2 resistance allele was significantly higher in non-infected (55.88%) compared to infected (40.99%) mosquitoes (Fisher's exact test, P<0.0001). Prevalence of infection was significantly higher in heterozygous and homozygous susceptible genotypes (P<0.001). However, homozygous resistant and heterozygous mosquitoes exhibited significantly higher infection intensity (P<0.01). No association was observed between the GSTe2 polymorphism and the infection status of mosquitoes.

Conclusion:Altogether, these results suggest that GSTe2-based metabolic resistance may affect the vectorial competence of resistantAn. funestusmosquitoes toP. falciparuminfection, by possibly increasing its permissiveness toPlasmodiuminfection.
An (Almost) Optimal Solution for Orthogonal Point Enclosure Query in ℝ3
Authors: Rahul Saladi
Published: October 2019

The orthogonal point enclosure query (OPEQ) problem is a fundamental problem in the context of data management for modeling user preferences. Formally, preprocess a set S of n axis-aligned boxes (possibly overlapping) in ℝd into a data structure so that the boxes in S containing a given query point q can be reported efficiently. In the pointer machine model, optimal solutions for the OPEQ in ℝ1 and ℝ2 were discovered in the 1980s: linear-space data structures that can answer the query in O(log n + k) query time, where k is the number of boxes reported. However, for the past three decades, an optimal solution in ℝ3 has been elusive. In this work, we obtain the first data structure that almost optimally solves the OPEQ in ℝ3 in the pointer machine model: an O(n log* n)-space data structure with O(log2 n · log log n + k) query time. Here, log* n is the iterated logarithm of n. This almost matches the lower-bound, which states that any data structure that occupies O(n) space requires Ω(log2 n + k) time to answer an OPEQ in ℝ3. Finally, we also obtain the best known bounds for the OPEQ in higher dimensions (d ≥ 4).
EDUCAÇÃO EM DIREITOS HUMANOS E AS QUESTÕES DE GÊNERO NA ESCOLA
Authors: Santos Ana Paula Da Silva , Brito Leandro Teófilo de
Published: October 2019

A questão dos direitos humanos tem se apresentado como uma problemática de difícil compreensão nos dias atuais, assim como o desafio do reconhecimento das diferenças. Mesmo com a construção de documentos nacionais e internacionais centrados na perspectiva dos direitos humanos, ainda convivemos com fortes violações, múltiplas violências, desigualdades sociais e diversas formas de preconceitos e discriminações no cotidiano das relações sociais.  Neste sentido, não podemos deixar de pensar nas questões que envolvem as relações de gênero e as desigualdades sofridas por meninas e mulheres, em diversos espaços sociais, sobretudo na escola. O presente estudo visa apresentar considerações sobre a educação em direitos humanos e as questões de gênero no contexto escolar, utilizando para tanto a perspectiva intercultural. Esta perspectiva nos coloca no âmbito de compreender a educação em direitos humanos como uma mediação para a construção de uma sociedade inclusiva, igualitária e plural onde os sujeitos possam vivenciar experiências que sejam permeadas pelos direitos humanos e, que, se tornem sujeitos de direito.