Extracellular Protease Inhibition Alters the Phenotype of Chondrogenically Differentiating Human Mesenchymal Stem Cells (MSCs) in 3D Collagen Microspheres

Matrix remodeling of cells is highly regulated by proteases and their inhibitors. Nevertheless, how would the chondrogenesis of mesenchymal stem cells (MSCs) be affected, when the balance of the matrix remodeling is disturbed by inhibiting matrix proteases, is incompletely known. Using a previously developed collagen microencapsulation platform, we investigated whether exposing chondrogenically differentiating MSCs to intracellular and extracellular protease inhibitors will affect the extracellular matrix remodeling and hence the outcomes of chondrogenesis. Results showed that inhibition of matrix proteases particularly the extracellular ones favors the phenotype of fibrocartilage rather than hyaline cartilage in chondrogenically differentiating hMSCs by upregulating type I collagen protein deposition and type II collagen gene expression without significantly altering the hypertrophic markers at gene level. This study suggests the potential of manipulating extracellular proteases to alter the outcomes of hMSC chondrogenesis, contributing to future development of differentiation protocols for fibrocartilage tissues for intervertebral disc and meniscus tissue engineering.published_or_final_versio

Meteorin regulates mesendoderm development by enhancing nodal expression

During gastrulation, distinct lineage specification into three germ layers, the mesoderm, endoderm and ectoderm, occurs through an elaborate harmony between signaling molecules along the embryonic proximo-distal and anterior-posterior axes, and Nodal signaling plays a key role in the early embryonic development governing embryonic axis formation, mesoderm and endoderm specification, and left-right asymmetry determination. However, the mechanism by which Nodal expression is regulated is largely unknown. Here, we show that Meteorin regulates Nodal expression and is required for mesendoderm development. It is highly expressed in the inner cell mass of blastocysts and further in the epiblast and extra-embryonic ectoderm during gastrulation. Genetic ablation of the Meteorin gene resulted in early embryonic lethality, presumably due to impaired lineage allocation and subsequent cell accumulation. Embryoid body culture using Meteorin-null embryonic stem (ES) cells showed reduced Nodal expression and concomitant impairment of mesendoderm specification. Meteorin-null embryos displayed reduced levels of Nodal transcripts before the gastrulation stage, and impaired expression of Goosecoid, a definitive endoderm marker, during gastrulation, while the proximo-distal and anterior-posterior axes and primitive streak formation were preserved. Our results show that Meteorin is a novel regulator of Nodal transcription and is required to maintain sufficient Nodal levels for endoderm formation, thereby providing new insights in the regulation of mesendoderm allocation.open1113sciescopu

Six SNPs and a TTG indel in sheep desmoglein 4 gene are in complete linkage disequilibrium

Desmoglein 4 (DSG4) plays an important role in the regulation of growth and differentiation of hair follicles in mammals. In this study, a 755 bp long segment of DSG4 was screened in 544 sheep sampled from nine Chinese indigenous breeds and two Western breeds using PCR-SSCP assay with three different pairs of primers. Two of the three fragments showed polymorphisms with genotypes defined as AA, AB, BB and BC, and DD, DE, and EE, respectively. Interestingly, polymorphisms in these two fragments were in strong linkage disequilibrium. Only three haplotypes were found, of which haplotype AD determined by alleles A and D was the major one in all breeds, while haplotype BE was only found in Chinese breeds that possess divergent frequencies ranging from 0.02 to 0.43; haplotype CD was very rare and present in only one Chinese sheep. Sequences of the three haplotypes showed seven single nucleotide polymorphisms (SNPs) and a TTG insertion/deletion (indel), leading to five amino acid substitutions and a glycine indel. Our study provides valuable genetic markers in evaluating the impact of the DSG4 gene on wool traits in sheep.Key words: Sheep, DSG4 gene, single-strand conformational polymorphism (SSCP), variation, linkage disequilibrium

Bio-Inspired Aggregation Control of Carbon Nanotubes for Ultra-Strong Composites

High performance nanocomposites require well dispersion and high alignment of the nanometer-sized components, at a high mass or volume fraction as well. However, the road towards such composite structure is severely hindered due to the easy aggregation of these nanometer-sized components. Here we demonstrate a big step to approach the ideal composite structure for carbon nanotube (CNT) where all the CNTs were highly packed, aligned, and unaggregated, with the impregnated polymers acting as interfacial adhesions and mortars to build up the composite structure. The strategy was based on a bio-inspired aggregation control to limit the CNT aggregation to be sub 20--50 nm, a dimension determined by the CNT growth. After being stretched with full structural relaxation in a multi-step way, the CNT/polymer (bismaleimide) composite yielded super-high tensile strengths up to 6.27--6.94 GPa, more than 100% higher than those of carbon fiber/epoxy composites, and toughnesses up to 117--192 MPa. We anticipate that the present study can be generalized for developing multifunctional and smart nanocomposites where all the surfaces of nanometer-sized components can take part in shear transfer of mechanical, thermal, and electrical signals

Gout penyakit lama dihidapi manusia

This paper proposes a differential evolution with local information for TSK-type neuro-fuzzy system optimization. The differential evolution with local information consider neighborhood between each individual to keep the diversity of population. An adaptive parameter tuning based on 1/5th rule is used to trade off between local search and global search. For structure learning algorithm, the on-line clustering algorithm is used for rule generation. The structure learning algorithm generates a new rule which compares the firing strength. Initially, there is no rule in neuro-fuzzy system model. The rules are automatically generated by fuzzy measure. For parameter learning, the parameters are optimized by differential evolution algorithm. Finally, the proposed neuro-fuzzy system with novel differential evolution model is applied in chaotic sequence prediction problem. Results of this paper demonstrate the effectiveness of the proposed model. © 2011 IEEE

Multidimensional Assessment of Asthma Identifies Clinically Relevant Phenotype Overlap: A Cross-Sectional Study.

BACKGROUND:Asthma is a heterogeneous disease with multiple phenotypes; however, the relevance of phenotype overlap remains largely unexplored. OBJECTIVE:To examine the relationship between phenotype overlap and clinical and inflammatory profiles of asthma. METHODS:In this cross-sectional study, adult participants with stable asthma (n = 522) underwent multidimensional assessments. The 10 most common phenotypes of asthma were defined and then classified into those commonly associated with Type (T) 2 or non-T2 inflammation. Furthermore, phenotype overlap scores (POS), representing the cumulative concomitant phenotypes, were used to analyze its association with clinical and inflammatory asthmatic profiles. RESULTS:Among the 522 participants, 73.4% (n = 383) had phenotype overlap, and mixed T2 and non-T2 inflammation coexisted in 47.5% (n = 248). T2 POS was positively associated with eosinophils, IgE, and fractional exhaled nitric oxide (FeNO), and negatively with Asthma Quality of Life Questionnaire (AQLQ), sputum neutrophils, IL-17A, IL-8, and TNF-α. Non-T2 POS was positively associated with Asthma Control Questionnaire, neutrophils and sputum IL-8, and negatively with AQLQ, forced expiratory volume in 1 s, blood eosinophils, IgE, and FeNO (all P < .05). Patients with phenotypes that are associated with mixed T2 and non-T2 inflammation had elevated T2 inflammation biomarkers but worse asthma control. Both T2 (adjusted β = -0.191, P = .035) and non-T2 (adjusted β = 0.310, P < .001) POS were significantly associated with severe exacerbations. CONCLUSIONS:Phenotype overlap is extremely common in asthmatic patients and significantly associated with clinical and inflammatory profiles. Patients with phenotypes associated with mixed T2 and non-T2 inflammation might be unresponsive to medications owing to increased non-T2 inflammation. Multidimensional asthma assessment identifies clinically relevant phenotype overlap

Anomalous tqγtq\gamma coupling effects in exclusive radiative B-meson decays

The top-quark FCNC processes will be searched for at the CERN LHC, which are correlated with the B-meson decays. In this paper, we study the effects of top-quark anomalous interactions $tq\gamma$ in the exclusive radiative $B\to K^*\gamma$ and $B\to\rho\gamma$ decays. With the current experimental data of the branching ratios, the direct CP and the isospin asymmetries, bounds on the coupling $\kappa_{tcR}^{\gamma}$ from $B\to K^*\gamma$ and $\kappa_{tuR}^{\gamma}$ from $B\to \rho\gamma$ decays are derived, respectively. The bound on $|\kappa_{tcR}^{\gamma}|$ from ${\mathcal B}(B\to K^{*}\gamma)$ is generally compatible with that from ${\mathcal B}(B\to X_{s}\gamma)$. However, the isospin asymmetry $\Delta(K^{*}\gamma)$ further restrict the phase of $\kappa_{tcR}^{\gamma}$, and the combined bound results in the upper limit, $\mathcal B(t\to c\gamma)<0.21$, which is lower than the CDF result. For real $\kappa_{tcR}^{\gamma}$, the upper bound on $\mathcal B(t\to c\gamma)$ is about of the same order as the $5\sigma$ discovery potential of ATLAS with an integrated luminosity of $10 {\rm fb}^{-1}$. For $B\to\rho\gamma$ decays, the NP contribution is enhanced by a large CKM factor $|V_{ud}/V_{td}|$, and the constraint on $tu\gamma$ coupling is rather restrictive, $\mathcal B(t\to u\gamma)<1.44\times 10^{-5}$. With refined measurements to be available at the LHCb and the future super-B factories, we can get close correlations between $B\to V \gamma$ and the rare $t\to q\gamma$ decays, which will be studied directly at the LHC ATLAS and CMS.Comment: 25 pages, 15 figures, pdflate

A novel tumor suppressor gene ECRG4 interacts directly with TMPRSS11A (ECRG1) to inhibit cancer cell growth in esophageal carcinoma

<p>Abstract</p> <p>Background</p> <p>The esophageal carcinoma related gene 4 (ECRG4) was initially identified and cloned from human normal esophageal epithelium in our laboratory (GenBank accession no.<ext-link ext-link-id="AF325503" ext-link-type="gen">AF325503</ext-link>). ECRG4 has been described as a novel tumor suppressor gene associated with prognosis in esophageal squamous cell carcinoma (ESCC).</p> <p>Methods</p> <p>In this study, binding affinity assay in vitro and co-immunoprecipitation experiment in vivo were utilized to verify the physical interaction between ECRG4 and transmembrane protease, serine 11A (TMPRSS11A, also known as ECRG1, GenBank accession no. <ext-link ext-link-id="AF 071882" ext-link-type="gen">AF 071882</ext-link>). Then, p21 protein expression, cell cycle and cell proliferation regulations were examined after ECRG4 and ECRG1 co-transfection in ESCC cells.</p> <p>Results</p> <p>We revealed for the first time that ECRG4 interacted directly with ECRG1 to inhibit cancer cell proliferation and induce cell cycle G1 phase block in ESCC. Binding affinity and co-immunoprecipitation assays demonstrated that ECRG4 interacted directly with ECRG1 in ESCC cells. Furthermore, the ECRG4 and ECRG1 co-expression remarkably upregulatd p21 protein level by Western blot (P < 0.001), induced cell cycle G1 phase block by flow cytometric analysis (P < 0.001) and suppressed cell proliferation by MTT and BrdU assay (both P < 0.01) in ESCC cells.</p> <p>Conclusions</p> <p>ECRG4 interacts directly with ECRG1 to upregulate p21 protein expression, induce cell cycle G1 phase block and inhibit cancer cells proliferation in ESCC.</p

Porphyromonas gingivalis suppresses adaptive immunity in periodontitis, atherosclerosis and Alzheimer’s disease

Porphyromonas gingivalis, a keystone pathogen in chronic periodontitis, has been found to associate with remote body organ inflammatory pathologies including atherosclerosis and Alzheimer’s disease (AD). Although P. gingivalis has a plethora of virulence factors, much of its pathogenicity is surprisingly related to the overall immunosuppression of the host. This review focuses on P. gingivalis aiding suppression of the host’s adaptive immune system involving manipulation of cellular immunological responses specifically T- and B-cells in periodontitis and related conditions. In periodontitis this bacterium inhibits the synthesis of IL-2 and increases humoral responses. This reduces inflammatory responses related to T- and B-cell activation, and subsequent IFN-ɤ secretion by a subset of T cells. The T cells further suppresses upregulation of programmed cell death-1 (PD-1)-receptor on CD+-cells and its ligand PD-L1 on CD11b+- subset of T-cells. IL-2 down-regulates immune response-regulated genes, induces a cytokine pattern in which the Th17 lineage is favored thereby modulating the Th17/ T-regulatory cell (Treg) imbalance. The suppression of IFN-ɤ stimulated release of interferon-inducible protein-10 (IP-10) chemokine ligands [ITAC (CXCL11) and Mig (CXCL9)] by P. gingivalis capsular serotypes, triggers distinct T-cell responses, and contributes to local immune evasion by release of its outer membrane vesicles. In atherosclerosis P. gingivalis reduces Tregs and transforming growth factor beta-1 (TGF-1) and causes imbalance in the Th17 lineage of the Treg population. In Alzheimer’s disease P. gingivalis may affect the blood-brain barrier permeability, and inhibit local IFN-ɤ response by preventing entry of immune cells into the brain. The scarcity of adaptive immune cells in Alzheimer’s disease neuropathology implies P. gingivalis infection of the brain likely causes impaired clearance of insoluble amyloid and induces immunosuppression. By the effective manipulation of the armory of adaptive immune suppression through a plethora of virulence factors P. gingivalis may act as a keystone organism in periodontitis and in related systemic diseases and other remote body inflammatory pathologies