Zingiber officinale Roscoe aqueous extract modulates Matrixmetalloproteinases and tissue inhibitors of Metalloproteinases expressions in Dengue virus-infected cells: implications for prevention of vascular permeability

Purpose: To investigate the effect of the aqueous extract of Zingiber officinale Roscoe. (ZOA) rhizome on the activity and expression of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 using an in vitro model of Dengue virus (DV) infection.Methods: Z. officinale rhizomes were extracted with water by continuous shaking for 5 days. The total phenolic content in extract was measured by Folin-Ciocalteu method. High performance liquid chromatography (HPLC) was employed to define qualitative and quantitative content of [6]-gingerol in ZOA. The median inhibitory concentration (IC50) value of ZOA for Vero cells was determined by 3-(4,5 dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide (MTT) assay. To induce MMPs production, Vero cells were infected with DV3. The modulatory effect of ZOA on the activity and expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed using gelatin zymography and quantitative Real-time polymerase chain reaction (RTPCR), respectively.Results: The yield of the ZOA was 7.98%. Total phenolics in ZOA was 68.17 ± 0.28 mg GAE/g of extract and it contained 29.32 ± 1.97 mg 6-gingerol/g of extract.The half-maximal inhibition concentration (IC50) of ZOA was 348.8 μg/mL for Vero cells. DV infection of Vero cells significantly elevated the production of soluble gelatinolytic MMP-2 and to a lesser extent, MMP-9, and their activities were significantly inhibited by ZOA in a dose-dependent manner. A significant down-regulation of MMP-2, MMP-9 mRNA expression and up-regulation of TIMP-1, TIMP-2 mRNA expression were observed in DV-infected Vero cells following treatment with ZOA, and it occurred in a  dose-dependent manner.Conclusion: The findings of this study suggest that ZOA may ameliorate plasma leakage in dengue virus infection and decrease the chances of severe dengue complications, dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) by inhibiting the activities and expression of MMP-2 and MMP-9 while upregulating the expression of TIMP-1 and TIMP-2

Caracterización y estudio de solubilidad de mezclas binarias de flunarizina / polietilenglicol 4000

En este trabajo estudiamos las interacciones de la flunarizina con polietilenglicol 4000 en dispersiones sólidas preparadassiguiendo el método de disolución propuesto por Sekiguchi y Obi. Como elementos de comparación se hanutilizado mezclas físicas de ambos componentes, preparados en las mismas proporciones de fármaco/polímero: 10/90, 20/80, 30/70, 40/60, 50/50,60/40 y 80/20.Las propiedades fisicoquímicas de las dispersiones sólidas y mezclas físicas se investigan mediante espectroscopíainfrarroja,difracción de rayos X, calorimetría diferencial de barrido y solubilidad en equilibrio. Los espectros deinfrarrojo indican que no ha habido interacción química entre la flunarizina y el PEG. Los difractogramas muestranque a determinadas proporciones, el PEG se introduce en la estructura de la flunarizina y los estudios térmicosparecen indicar la formación de una mezcla eutéctica a la proporción 28,96% de flunarizina y 71,04% de PEG 4000.Todas las muestras presentan una solubilidad superior a la del fármaco puro y en ambos tipos de muestras elincremento es mayor al aumentar la proporción de polímero. El análisis de comparación múltiple aplicado independientementea las dispersiones sólidas y mezclas físicas, indica que no existe diferencia estadísticamente significativa(p<0,05) entre las muestras de proporciones 30/70, 40/60, 50/50 y 60/40, pero sí hay diferencias entre éstas y las deproporciones 10/90, 20/80 y 80/20

Effects of Combining Feed Grade Urea and a Slow-release Urea Product on Performance, Dietary Energetics and Carcass Characteristics of Feedlot Lambs Fed Finishing Diets with Different Starch to Acid Detergent Fiber Ratios.

Recent findings have shown that microbial nitrogen flow and digestible energy of diets are increased when urea is combined with a slow-release urea (SRU) in diets with a starch to acid detergent fibre ratio (S:F) 4:1. This affect is attributable to enhanced synchrony between ruminal N availability for microbial growth and carbohydrate degradation. To verify the magnitude of this effects on lamb performance, an experiment was conducted to evaluate the effects of combining urea and a SRU in diets containing S:F ratios of 3:1, 4:1, or 5:1 on performance, dietary energetics and carcass characteristics of finishing lambs. For that, 40 Pelibuey×Katahdin lambs (36.65±3 kg) were assigned to one of five weight groupings in 20 pens (5 repetition/treatments). The S:F ratio in the diet was manipulated by partially replacing the corn grain and dried distiller's grain with solubles by forage (wheat straw) and soybean meal to reach S:F ratios of 3:1, 4:1 or 5:1. An additional treatment of 4:1 S:F ratio with 0.8% urea as the sole source of non-protein nitrogen was used as a reference for comparing the effect of urea combination vs. conventional urea at the same S:F ratio. There were no treatment effects on dry matter intake (DMI). Compared the urea combination vs urea at the same S:F ratio, urea combination increased (p<0.01) average daily gain (ADG, 18.3%), gain for feed (G:F, 9.5%), and apparent energy retention per unit DMI (8.2%). Irrespective of the S:F ratio, the urea combination improved the observed-to-expected dietary ratio and apparent retention per unit DMI was maximal (quadratic effect, p≤0.03) at an S:F ratio of 4:1, while the conventional urea treatment did not modify the observed-to-expected net energy ratio nor the apparent retention per unit DMI at 4:1 S:F ratio. Urea combination group tended (3.8%, p = 0.08) to have heavier carcasses with no effects on the rest of carcass characteristics. As S:F ratio increased, ADG, G:F, dietary net energy, carcass weight, dressing percentage and longissimus thoracis (LM) area increased linearly (p≤0.02). Combining urea and a slow-release urea product results in positive effects on growth performance and dietary energetics, but the best responses are apparently observed when there is a certain proportion (S:F ratio = 4:1) of starch to acid detergent fibre in the diet

Percutaneous Closure of Atrial Septal Defects: a Decade of Experience at a Reference Center

INTRODUCTION: Atrial septal defects (ASD) are among the most common congenital anomalies and account for 10% of congenital heart disease in the pediatric age-group and 30% in adults. Closure is indicated when there is evidence of hemodynamic significance or after a paradoxical embolic event. Ten years ago, percutaneous closure became the treatment of choice in our center for all patients with a clear indication and favorable anatomy. In this paper we report the experience of this first decade. OBJECTIVE: To assess the short- and long-term results of our ten-year experience with percutaneous closure of atrial septal defects. METHODS: We studied retrospectively all patients with ASD treated with a percutaneous approach between November 1998 and December 2008. The pediatric age-group consisted of patients younger than 19 years old. Demographic data, clinical indications, minor and major complication rates, success rate and long-term outcome were assessed. RESULTS: In the first ten years of experience 510 patients, of whom 166 were in the pediatric group, were treated in our center by a team of adult and pediatric cardiologists. The overall success rate of the procedure was 98% (97.5% in ASD and 99.5% in patent foramen ovale (PFO). The minor complication rate was 3% (3.4% in ASD and 2% in PFO). The most frequent complication was supraventricular tachycardia. The major complication rate was 1.2% (0.6% in ASD and 2% in PFO). Two patients developed cardiac tamponade due to hemopericardium that was resolved by pericardiocentesis, without need for surgery. One patient had an arterial pseudoaneurysm corrected by vascular surgery. There was no device embolization and no need for urgent surgery in this population. During follow-up two patients had recurrence of ischemic stroke, one had a transient ischemic attack and another had a hemorrhagic stroke. Mortality was 0.6% (0.6% in ASD and 0.5% in PFO). There were no in-hospital deaths. During follow-up there were two deaths, both in the adult group. DISCUSSION AND CONCLUSION: In this population the success rate was high and most of the complications were minor. The results of this collaboration between adult and pediatric cardiologists in the first ten years of activity confirm the safety and efficacy of percutaneous closure of septal defects, when there is careful patient selection and a standardized technique

Safeguarding children in dentistry: 1. Child protection training, experience and practice of dental professionals with an interest in paediatric dentistry

* Few dental professionals with child protection training have experience of making referrals. * There is a wide gap in practice between recognising signs of child abuse and neglect and responding effectively. * This may indicate missed opportunities to save children from continuing abuse. * There is a need for improved child protection information, support and training for dental professionals. Abstract Following several highly publicised inquiries into the deaths of children from abuse and neglect, there has been much recent interest in the role and responsibility of all health professionals to protect children at risk of maltreatment. The findings of a postal questionnaire, sent in March 2005 to 789 dentists and dental care professionals with an interest in paediatric dentistry working in varied settings in the UK, are presented in a two-part report and discussed in the context of current multi-agency good practice in safeguarding and promoting the welfare of children. This first part explores reported child protection training, experience and practice. There was a significant gap between recognising signs of abuse and responding effectively: 67% of respondents had suspected abuse or neglect of a child patient at some time in their career but only 29% had ever made a child protection referral. The dental profession is alerted to the need to ensure necessary appropriate action to safeguard children is always taken when child abuse or neglect are suspected

Does native Trypanosoma cruzi calreticulin mediate growth inhibition of a mammary tumor during infection?

Indexación: Web of Science.Background: For several decades now an antagonism between Trypanosoma cruzi infection and tumor development has been detected. The molecular basis of this phenomenon remained basically unknown until our proposal that T. cruzi Calreticulin (TcCRT), an endoplasmic reticulum-resident chaperone, translocated-externalized by the parasite, may mediate at least an important part of this effect. Thus, recombinant TcCRT (rTcCRT) has important in vivo antiangiogenic and antitumor activities. However, the relevant question whether the in vivo antitumor effect of T. cruzi infection is indeed mediated by the native chaperone (nTcCRT), remains open. Herein, by using specific modified anti-rTcCRT antibodies (Abs), we have neutralized the antitumor activity of T. cruzi infection and extracts thereof, thus identifying nTcCRT as a valid mediator of this effect. Methods: Polyclonal anti-rTcCRT F(ab')(2) Ab fragments were used to reverse the capacity of rTcCRT to inhibit EAhy926 endothelial cell (EC) proliferation, as detected by BrdU uptake. Using these F(ab')(2) fragments, we also challenged the capacity of nTcCRT, during T. cruzi infection, to inhibit the growth of an aggressive mammary adenocarcinoma cell line (TA3-MTXR) in mice. Moreover, we determined the capacity of anti-rTcCRT Abs to reverse the antitumor effect of an epimastigote extract (EE). Finally, the effects of these treatments on tumor histology were evaluated. Results: The rTcCRT capacity to inhibit ECs proliferation was reversed by anti-rTcCRT F(ab')(2) Ab fragments, thus defining them as valid probes to interfere in vivo with this important TcCRT function. Consequently, during infection, these Ab fragments also reversed the in vivo experimental mammary tumor growth. Moreover, anti-rTcCRT Abs also neutralized the antitumor effect of an EE, again identifying the chaperone protein as an important mediator of this anti mammary tumor effect. Finally, as determined by conventional histological parameters, in infected animals and in those treated with EE, less invasive tumors were observed while, as expected, treatment with F(ab')(2) Ab fragments increased malignancy. Conclusion: We have identified translocated/externalized nTcCRT as responsible for at least an important part of the anti mammary tumor effect of the chaperone observed during experimental infections with T. cruzi.http://bmccancer.biomedcentral.com/articles/10.1186/s12885-016-2764-

The gradient flow running coupling with twisted boundary conditions

We study the gradient flow for Yang-Mills theories with twisted boundary conditions. The perturbative behavior of the energy density $\langle E(t)\rangle$ is used to define a running coupling at a scale given by the linear size of the finite volume box. We compute the non-perturbative running of the pure gauge $SU(2)$ coupling constant and conclude that the technique is well suited for further applications due to the relatively mild cutoff effects of the step scaling function and the high numerical precision that can be achieved in lattice simulations. We also comment on the inclusion of matter fields.Comment: 27 pages. LaTe

The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster

The regulatory protein NsrR, a member of the Rrf2 family of transcription repressors, is specifically dedicated to sensing nitric oxide (NO) in a variety of pathogenic and non-pathogenic bacteria. It has been proposed that NO directly modulates NsrR activity by interacting with a predicted [Fe-S] cluster in the NsrR protein, but no experimental evidence has been published to support this hypothesis. Here we report the purification of NsrR from the obligate aerobe Streptomyces coelicolor. We demonstrate using UV-visible, near UV CD and EPR spectroscopy that the protein contains an NO-sensitive [2Fe-2S] cluster when purified from E. coli. Upon exposure of NsrR to NO, the cluster is nitrosylated, which results in the loss of DNA binding activity as detected by bandshift assays. Removal of the [2Fe-2S] cluster to generate apo-NsrR also resulted in loss of DNA binding activity. This is the first demonstration that NsrR contains an NO-sensitive [2Fe-2S] cluster that is required for DNA binding activity