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The study was aimed to analyse the effects of miR-128 on the proliferation and apoptosis of lung cancer cells of non-small cell by regulating RECK. The expression of miR-128 in non-small cell lung cancer (NSCLC) and its paracancerours tissues was analyzed by sequencing. miR-128 expression level in NSCLC and its paracancerours tissues was detected by qPCR. We over expressed miR-128 by mimics in A549 NSCLC cell line. Establish blank and negative control group. miR-128 and RECK expression level in each group was detected by qPCR. Use MTT assay to test the proliferation ability of each group. The apoptosis level and the level of ROS in each group were tested by hoechst 33258. The expression level of apoptosis-related protein and p38 NF- B signal pathway-related protein in each group was tested by Western blot. The results of sequencing and qPCR showed that compared with the blank control group, the expression level of miR-128 mRNA was significantly higher in the adjacent tissues than in the adjacent tissues (P < 0 05). The expression levels of miR128 mRNA and RECK mRNA in the overexpression group were significantly increased (P

In this paper, we explored a convenient method for preparing stable silver nanoclusters (AgNCs) within 1 h at room temperature using keratin as a template and NaBH4 as a reducing agent. As a group of cysteine-rich structural proteins, keratins are abundant in wool, hair and feathers. The prepared AgNCs exhibited high fluorescence intensity and stable emission at 705 nm. Their fluorescence intensity decreased by

Background The circulating levels of Cyr61 (also known as CCN1) may prove to have great clinical value in the diagnosis, monitoring and prognosis of many disorders in humans. However, the reference intervals (RIs) for this analyte in human subjects have not previously been well established. Therefore, establishing RIs and determining the distribution of circulating Cyr61 levels are very important for future clinical studies and could provide an orientation value for exploring its clinical usefulness. Methods The Cyr61 levels in 2,514 healthy Chinese Han subjects (1,250 males and 1,264 females, aged 18 - 88 years, recruited from 4 hospitals in Shanghai and Fujian) were measured with a sandwich ELISA (R&D Systems, USA). The RIs were determined in a manner consistent with the Clinical and Laboratory Standards Institute guidelines. Results The levels of serum Cyr61 showed a non-Gaussian distribution. A statistically significant difference was observed between the males and females such that the median level of Cyr61 in the males was significantly higher than that in the females. Furthermore, the Cyr61 levels significantly increased with age in the female group whereas no difference was observed among the different age groups among the males. The RIs for serum Cyr61 were 3.3 - 184 pg/mL and 5.0 - 182 pg/mL in females aged 18 - 45 and 46 - 88 years, respectively. The RI for serum Cyr61 was 4.0 - 198 pg/mL in the males. Conclusions The RIs for serum Cyr61 were established among Chinese Han individuals. The effects of age and gender on the distribution characteristics of serum Cyr61 were studied, revealing that the RIs were gender and, in females, age-specific, which may suggest that a female hormone, estrogen plays a role in the regulation of Cyr61 expression in vivo.

Metallothionein (MT), a cysteine rich protein is involved as a radical scavenger in several pathological conditions associated with oxidative stress; however, its role in periodontal disease still remains elusive. The aim of this cross-sectional study is to determine the serum, saliva and gingival crevicular fluid (GCF) levels of MT in smokers (S) and non-smokers (NS) with chronic periodontitis (CP), and compare them with those of periodontally healthy (PH) individuals.A total of 85 participants were enrolled: 45 patients with CP (23 S [CP+S] and 22 NS [CP+NS]) and 40 PH individuals (20 S [PH+S] and 20 NS [PH+NS]). In all the study participants, clinical periodontal parameters (plaque index, gingival index, sulcus bleeding index, probing depth, and clinical attachment level) were recorded and samples of serum, saliva and GCF were collected. Enzyme-linked immunosorbent assay was used to determine the levels of MT in the samples.All periodontal clinical parameters were significantly higher in the CP groups as compared to PH groups (P 0.05). MT levels in CP+S group were significantly raised in comparison to other three groups. There was no statistically significant difference in MT levels among CP+NS and PH+S groups (P 0.05); however, relatively higher levels were observed in GCF and saliva in CP+NS group. When all the study groups were observed together, MT levels were positively correlated with clinical parameters.Results of present study suggest that smoking and CP can induce the synthesis of MT owing to increased oxidative stress and heavy metals intoxication. Further longitudinal studies with large sample size and an interventional arm are needed to substantiate the role of MT as a potential biomarker in periodontitis.

Aim: The protein CCN1/CYR61 exerts critical functions in myocardial ischemic injury. We sought to investigate the prognostic value of CCN1 in patients with acute heart failure (AHF) and coronary heart disease (CAD). Methodology: We prospectively enrolled 113 patients with AHF and CAD. Patients were followed for all-cause mortality during a 30-day follow-up. Logistic models were used to estimate the association of CCN1 concentrations with 30-day mortality. Results: In multivariate logistic regression model, CCN1 was a significant predictor of 30-day mortality independent of current markers. Enhanced Feedback for Effective Cardiac Treatment risk score was recommended as one of the selected multivariable risk scores to predict outcome in AHF. CCN1 improved risk stratification for all-cause mortality when added to the Enhanced Feedback for Effective Cardiac Treatment risk scores at 30 days. Conclusion: We found CCN1 is independently associated with 30-day mortality in patients with AHF and CAD.

Clostridium difficile is a Gram-positive spore-former bacterium and the leading cause of nosocomial antibiotic-associated diarrhea that can culminate in fatal colitis. During the infection, C. difficile produces metabolically dormant spores, which persist in the host and can cause recurrence of the infection. The surface of C. difficile spores seems to be the key in spore-host interactions and persistence. The proteome of the outermost exosporium layer of C. difficile spores has been determined, identifying two cysteine-rich exosporium proteins, CdeC and CdeM. In this work, we explore the contribution of both cysteine-rich proteins in exosporium integrity, spore biology and pathogenesis. Using targeted mutagenesis coupled with transmission electron microscopy we demonstrate that both cysteine rich proteins, CdeC and CdeM, are morphogenetic factors of the exosporium layer of C. difficile spores. Notably, cdeC, but not cdeM spores, exhibited defective spore coat, and were more sensitive to ethanol, heat and phagocytic cells. In a healthy colonic mucosa (mouse ileal loop assay), cdeC and cdeM spore adherence was lower than that of wild-type spores; while in a mouse model of recurrence of the disease, cdeC mutant exhibited an increased infection and persistence during recurrence. In a competitive infection mouse model, cdeC mutant had increased fitness over wild-type. Through complementation analysis with FLAG fusion of known exosporium and coat proteins, we demonstrate that CdeC and CdeM are required for the recruitment of several exosporium proteins to the surface of C. difficile spores. CdeC appears to be conserved exclusively in related Peptostreptococcaeace family members, while CdeM is unique to C. difficile. Our results sheds light on how CdeC and CdeM affect the biology of C. difficile spores and the assembly of the exosporium layer and, demonstrate that CdeC affect C. difficile pathogenesis., Author summary We discovered a mechanism of assembly of the outer most layer of Clostridium difficile spores, the exosporium. While CdeC is conserved in several Peptostreptococcaeace family members, CdeM is unique to C. difficile. We show that two proteins that are rich in cysteine amino acid residues, CdeC and CdeM, are essential for the recruitment of additional spore coat and exosporium proteins. The absence of CdeC, had profound implications in the correct spore coat assembly which were related to decreased spore resistant properties that are relevant for in vivo infection such as lysozyme resistance, macrophage infection. Notably, the absence of either cysteine rich proteins leads to a decrease in spore adherence of C. difficile spores to healthy colonic mucosa; but only the absence of CdeC affected in vivo competitive fitness in a mouse model, recurrence of the disease in a mouse model of recurrent infection. Considering the importance of the outer layers of C. difficile spores in spore-host interactions, our findings have broad implications on the biology of C. difficile spores and to C. difficile pathogenesis.

Aim: The aim is to evaluate serum cysteine-rich protein 61 (Cyr61) levels in patients with coronary artery disease (CAD). Patients & methods: Serum Cyr61 levels were measured in 180 patients with CAD and 74 participants without CAD. Results: Serum Cyr61 levels were significantly higher in CAD patients. Patients with acute coronary syndrome showed significantly higher Cyr61 than those with stable angina pectoris. Serum Cyr61 levels in complex lesion group were significantly higher. Serum Cyr61 was positively correlated with Gensini score and C-reactive protein. Multivariable logistic regression analyses demonstrated that serum Cyr61 levels were independently correlated with the existence of CAD (p = 0.01). Conclusion: Our study suggested Cyr61 as a potential biomarker in characterizing CAD and therapeutic target for CAD.

In spite of the development of treatments, long-term survival in patients with glioblastoma is very rare. Recently, studies about clarifying prognostic markers for glioblastoma have been growing. Previously we reported that median progression-free survival and overall survival (OS) of patients with high cysteine-rich protein 61 (CCN1) expression were significantly shorter than those of patients with low CCN1 expression. To investigate the molecular mechanisms that regulate CCN1 expression, we analysed 147 tumor samples from 80 glioblastoma patients and 67 lower grade glioma patients using next-generation and Sanger sequencing. Sequencing analysis showed that CCN1 high expression group includes PIK3R1Met326Ile more frequently (10/37 cases, 27.0%) than CCN1 low expression group (3/38 cases, 7.9%) in glioblastoma. We also found this mutation in corresponding blood samples. And multivariate analysis indicated that, high CCN1 expression and PIK3R1Met326Ile in glioblastoma patients were prognostic factors for OS ([HR=2.488 (1.298–4.769), p=0.006] and [HR=2.089 (1.020–4.277), p=0.0439], respectively. According to our results, the PIK3R1Met326Ile mutation related to CCN1 expression and poor prognosis in glioblastoma.

Cell-cell communication is pivotal for the coordination of various features of plant development. Recent studies in plants have revealed that, as in animals, secreted signal peptides play critical roles during reproduction. However, the precise signalling mechanisms in plants are not well understood. In this review, we discuss the known and putative roles of secreted peptides present in the seeds of angiosperms as key signalling factors involved in coordinating different aspects of seed development.

Periodontal diseases are characterized by pathological destruction of extracellular matrix (ECM) of periodontal tissues. Matrix metalloproteinases (MMPs) are a significant part of the degradation of ECM. However, the regulation of MMPs expression level in periodontal diseases is as yet undetermined. RECK (reversion-inducing cysteine-rich protein with Kazal motifs), a novel membrane-anchored inhibitor of MMPs, could regulate the expressions of MMP-2, 9 and MT1-MMP as a cell surface-signaling molecule. Thus, we propose that RECK may play an important role in regulating MMPs in the ECM degradation of periodontal diseases. The RECK/MMPs signaling pathway could provide a new approach for prevention and treatment of RECK in periodontal diseases by blocking MMPs.

Despite advances in treatment, glioblastoma remains a lethal brain tumor. Efforts to identify prognostic molecular biomarkers for glioblastoma have intensified, and we previously reported that the median progression-free survival (PFS) and overall survival (OS) of patients with high levels of cysteine-rich protein 61 (CCN1/CYR61) are significantly shorter compared with those with low levels of CCN1. To identify the molecular mechanisms that regulate the expression of CCN1 in glioblastomas, we analyzed tumor samples from 42 patients with glioblastoma. Next-generation and Sanger sequencing techniques revealed that the PIK3R1Met326Ile mutation was significantly more frequent in patients with high vs low levels of CCN1 (9/22, 40.9% vs 2/20, 10.0%; respectively). Moreover, this mutation was identified in the corresponding blood samples. Univariate analysis of Kaplan-Meier survival data revealed that high levels of CCN1 and the presence of the PIK3R1Met326Ile mutation were significantly associated with PFS (p=0.0003 and 0.0191, respectively) and that age, high levels of CCN1, and the presence of the PIK3R1Met326Ile mutation were significantly associated with OS (p=0.0016, 0.0019 and 0.0057, respectively). Multivariate analysis identified age >65 years and the presence of the PIK3R1Met326Ile mutation as prognostic factors for OS (HR=3.533 [1.519-8.220], p=0.003) and (HR=2.712 [1.187-6.198], p=0.018, respectively). We conclude that the germline mutation PIK3R1Met326Ile correlates with the levels of CCN1 and poor prognosis of glioblastoma.

The complete nucleotide sequence of a sweet potato virus, first identified two decades ago as virus "C-6", was determined in this study. Sequence similarity and phylogenetic analysis clearly place it as a member of a distinct species within the genus Carlavirus, family Betaflexiviridae. Its genome structure was typical for that of other carlaviruses except that the ORF for the cysteine-rich protein was replaced by an ORF encoding a predicted protein with no similarity to any known protein.

Microbubbles have the potential to be used in many applications in different fields, especially in food, biomedical, and environmental sciences. However, to produce microbubbles with relatively long-term stability is difficult as they are inherently unstable entities. Hydrophobins are naturally occurring proteins that have previously been shown to have an ability to stabilize micron-sized air cells. More recently, small air cells with other cysteine-rich proteins such as bovine serum albumin and egg albumen (egg white protein) have been constructed using a sonochemical method. The majority of the air cells had a diameter between 1 and 10 μm. A dependence of the air cell size and stability with the processing conditions and concentration were found. The air cells’ shells are dense enough to form a cage-like structure around air cells and are equally as robust as those from hydrophobins.

When developing low-fat foods, one of the main concerns is the sensory performance which is generally inferior to that of the full-fat version. Micron-sized air cells coated and stabilised by proteins (0.5―10 μm) have been produced sonochemically using different cysteine-rich proteins (hydrophobin, bovine serum albumin [BSA] and egg white proteins [EWPs]). These suspensions of air cells have been termed "air filled emulsions" (AFEs) and suggested for the production of low-fat emulsion-based products. This study explores the oral (tribological) behavior of AFEs as ingredients and within a triphasic A/O/W emulsion-based prototype salad dressing. Tribological measurement of AFEs yielded different results for BSA-AFE and EWP-AFE, indicating that the very nature of the protein may play a crucial role. However, the triphasic A/O/W emulsion showed similar, if not better, lubrication properties than the standard O/W version, indicating that AFEs may contribute to the perceived fat-related attributes.

Smooth muscle-specific transcription is controlled by a multitude of transcriptional regulators that cooperate to drive expression in a temporospatial manner. Previous analysis of the cysteine-rich protein 1 ( CRP1/Csrp) gene revealed an intronic enhancer that is sufficient for expression in arterial smooth muscle cells and requires a serum response factor-binding CArG element for activity. The presence of a CArG box in smooth muscle regulatory regions is practically invariant; however, it stands to reason that additional elements contribute to the modulation of transcription in concert with the CArG. Because of the potential importance of other regulatory elements for expression of the CRP1 gene, we sought to identify additional motifs within the enhancer that are necessary for expression. In this effort, we identified a conserved cAMP response element (CRE) that, when mutated, diminishes the expression of the enhancer in cultured vascular smooth muscle cells. Using transfection and electrophoretic mobility shift assays, we have shown that the CRE binds the cAMP response element-binding protein (CREB) and is activated by Ca2+/calmodulin-dependent protein kinase IV (CaMKIV), but not by CaMKII. Furthermore, our data demonstrate that CaMKIV stimulates CRP1 expression not only through the CRE but also through the CArG box. These findings represent evidence of a functional CRE within a smooth muscle-specific gene and provide support for a mechanism in which CREB functions as a smooth muscle determinant through CaMKIV activation.

Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a novel membrane-anchored matrix metalloproteinase inhibitor, and experimental studies have shown that RECK can suppress tumor progression through angiogenesis inhibition. We have already revealed that enhanced RECK expression is significantly correlated with a favorable prognosis in non-small-cell lung cancer (NSCLC). In this study, further analyses focused on pN2 disease were conducted to assess the clinical significance of RECK expression.A total of 118 patients with completely resected pathologic stage IIIA N2 NSCLC were retrospectively examined. RECK expression in the primary tumor, along with involved N2 nodes, was examined immunohistochemically.RECK expression in the primary tumor was strong in 53 patients (44.9%) and was weak in the other 65 patients. The 5-year survival rate of patients with RECK-strong tumor (42.9%) was significantly higher than that of patients with RECK-weak tumor (23.1%; P = .017). Reduced RECK expression significantly correlated with a poor prognosis for patients with a single N2 node involved (P = .019), but not for patients with multiple N2 nodes involved (P = .440). A multivariate analysis confirmed that reduced RECK expression was an independent and significant factor to predict a poor prognosis (P = .031). RECK expression in involved N2 nodes was significantly higher than in primary tumors (P.001).RECK status was a novel prognostic factor in pathologic stage IIIA N2 NSCLC.

Emerging data have suggested that acute kidney injury (AKI) is often incompletely repaired and can lead to chronic kidney disease (CKD), which is characterized by tubulointerstitial inflammation and fibrosis. However, the underlying mechanisms linking AKI to CKD remain obscure. The present study aimed to investigate the role of cysteine-rich protein 61 (Cyr61) after unilateral kidney ischemia-reperfusion injury (IRI) in mice. After IRI, increased expression of Cyr61 was detected, predominately in the proximal tubular epithelium. This was confirmed by in vitro experiments, which showed that hypoxia stimulated Cyr61 expression in cultured proximal tubular epithelial cells. The proinflammatory property of Cyr61 was indicated by its ability to upregulate monocyte chemoattractant protein-1 and IL-6. Additionally, we found elevated urinary Cyr61 excretion in patients with AKI. Notably, treatment of mice with an anti-Cyr61 antibody attenuated the upregulation of kidney monocyte chemoattractant protein-1, IL-6, IL-1β, and macrophage inflammatory protein-2 and reduced the infiltration of F4/80-positive macrophages on days 7 and 14 after IRI. In addition, blockade of Cyr61 reduced the mRNA expression of collagen, transforming growth factor-β, and plasminogen activator inhibitor-I as well as the degree of collagen fibril accumulation, as evaluated by picrosirius red staining, and levels of α-smooth muscle actin proteins by day 14. Concurrently, in the treated group, peritubular microvascular density was more preserved on day 14. We conclude that Cyr61 blockade inhibits the triad of inflammation, interstitial fibrosis, and capillary rarefaction after severe ischemic AKI. The results of this study expand the knowledge of the mechanisms underlying the AKI-to-CKD transition and suggest that Cyr61 is a potential therapeutic target.

The mechanism by which the contractile machinery of muscle is assembled and maintained is not well-understood. Members of the cysteine-rich protein (CRP) family have been implicated in these processes. Three vertebrate CRPs (CRP1-3) that exhibit developmentally regulated muscle-specific expression have been identified. All three proteins are associated with the actin cytoskeleton, and one has been shown to be required for striated muscle structure and function. The vertebrate CRPs identified to date display a similar molecular architecture; each protein is comprised of two tandemly arrayed LIM domains, protein-binding motifs found in a number of proteins with roles in cell differentiation. Each LIM domain coordinates two Zn(II) ions that are bound independently in CCHC (C=Cys, H=His) and CCCC modules. Here we describe the solution structure of chicken CRP1 determined by homonuclear and 1H-15N heteronuclear magnetic resonance spectroscopy. Comparison of the structures of the two LIM domains of CRP1 reveals a high degree of similarity in their tertiary folds. In addition, the two component LIM domains represent two completely independent folding units and exhibit no apparent interactions with each other. The structural independence and spatial separation of the two LIM domains of CRP1 are compatible with an adapter or linker role for the protein.

The family of cysteine rich proteins (CRP) comprises three closely homologous members that have been reported to interact with α-actinin. Muscular LIM protein (MLP/CRP3), the skeletal muscle variant, was originally discovered as a positive regulator of myogenesis and is suggested to be part of the stretch sensor of the myofibril through its interaction with telethonin (T-Cap). We determined the structure of both LIM domains of human MLP by nuclear magnetic resonance spectroscopy. We confirm by 15N relaxation measurements that both LIM domains act as independent units and that the adjacent linker regions are fully flexible. With the published structures of CRP1 and CRP2, the complete family has now been structurally characterized.

International audience; Plant lipid transfer proteins are small soluble extracellular proteins that are able to bind and transfer a variety of lipids in vitro. Recently, it has been proposed that lipid transfer proteins may play a key role in plant defence mechanisms, especially during the induction of systemic acquired resistance. However, very little is known about the proteins expressed in developing plants and tissues, since almost all the biophysical and structural data available to date on lipid transfer proteins originate from proteins present in storage tissues of monocot cereal seeds. In this paper, we report the structural and functional characteristics of a lipid transfer protein (named LTP1_1) constitutively expressed in young aerial organs of Nicotiana tabacum (common tobacco). The unlabelled and uniformly labelled proteins were produced in the yeast Pichia pastoris, and we determined the three-dimensional (3D) structure of LTP1_1 using nuclear magnetic resonance (NMR) spectroscopy and molecular modeling techniques. The global fold of LTP1_1 is very close to the previously published structures of LTP1 extracted from cereal seeds, including an internal cavity. However, the chemical shift variations of several NMR signals upon lipid binding show that tobacco LTP1_1 is able to bind only one LysoMyristoylPhosphatidylCholine (LMPC), while wheat and maize LTPs can bind either one or two. Titration experiments using intrinsic tyrosine fluorescence confirm this result not only with LMPC but also with two fatty acids. These differences can be explained by the presence in tobacco LTP1_1 of a hydrophobic cluster closing the second possible access to the protein cavity. This result suggests that LTP1 lipid binding properties could be modulated by subtle changes in a conserved global structure. The biological significance of this finding is discussed in the light of the signalling properties of the tobacco LTP1_1-jasmonate complex described elsewhere.

on final consolidation are achieved with daily administration of 40 μg/kg of PTH1-34. Our results suggest the existence of a natural window of treatment efficacy in mouse peaking 18 d. after fracture. Healing processes are anticipated and enhanced from early teriparatide administration. Further applications of the study could be the treatment of osteoporotic or secondary fractures, consolidation delays and non-unions. Disclosure of Interest: None declared

Recent progress in molecular engineering has contributed to the great progress of medicine. However, there are still difficult problems constituting a challenge for molecular biology and biotechnology, e.g. new generation of anticancer agents, alternative biosensors or vaccines. As a biotechnological tool, bacteriophages (phages) offer a promising alternative to traditional approaches. They can be applied as anticancer agents, novel platforms in vaccine design, or as target carriers in drug discovery. Phages also offer solutions for modern cell imaging, biosensor construction or food pathogen detection. Here we present a review of bacteriophage research as a dynamically developing field with promising prospects for further development of medicine and biotechnology.

Background Cysteine rich protein families are notoriously difficult to align due to low sequence identity and frequent insertions and deletions. Results Here we present an alignment method that ensures homologous cysteines align by assigning a unique 10 amino acid barcode to those identified as structurally homologous by the DALI webserver. The free inter-cysteine regions of the barcoded sequences can then be aligned using any standard algorithm. Finally the barcodes are replaced with the original columns to yield an alignment which requires the minimum of manual refinement. Conclusions Using structural homology information to constrain sequence alignments allows the alignment of highly divergent, repetitive sequences that are poorly dealt with by existing algorithms. Tools are provided to perform this method online using the CysBar web-tool (http://CysBar.science.latrobe.edu.au) and offline (python script available from http://github.com/ts404/CysBar). Electronic supplementary material The online version of this article (doi:10.1186/s40064-015-1609-z) contains supplementary material, which is available to authorized users.