Antimicrobial synergism against different lineages of methicillin-resistant Staphylococcus aureus carrying SCCmec IV

Aim To evaluate the synergistic activity of antimicrobial drugs against lineages of methicillin-resistant Staphylococcus aureus (MRSA) carrying SCCmec IV. The biofilm production and related genes were also detected. Methods and Results Forty two MRSA isolates were tested for biofilm production and related genes. Biofilm/biomass susceptibility to gentamicin (G), linezolid (L), rifampicin (R) and vancomycin (V) was determined for six isolates from three lineages prevalent in Rio de Janeiro hospitals in concentrations ranging from 0·25 to 64 μg ml−1. Biomass was evaluated by microtitre plate test and number of viable cells (CFU cm−2) and inspected by epifluorescence microscopy. All isolates presented the icaA and sasG genes, but only 38% were biofilm producers. There were 50 and 45% biomass reductions when concentrations ≥4 μg ml−1 of R or L and ≥16 μg ml−1 of G or V, respectively, were used. Synergism tests produced a 55% biomass reduction with R2lg ml1 + G16lg ml1 , R2lg ml1 + L2lg ml1 , R2lg ml1 + V4lg ml1 , and L2lg ml1 + V4lg ml1 . Number of viable cells was reduced from 2 to 3 logs with R2lg ml1 + L2lg ml1 and R2lg ml1 + V4lg ml1 . Conclusions Synergisms involving R plus L and R plus V caused important reductions in biofilm/biomass and the number of viable cells. Drug combinations should be considered in the chemotherapies of MRSA-SCCmec IV infections. Significance and Impact of the Study Biofilms in MRSA infections restrict the clinical choice of antimicrobials. Thus, knowledge of the best options for monotherapy and drug synergisms could improve clinical results.This study was supported by grants from Fundacao Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), Coordenacao de Aperfeicoamento Pessoal de Nivel Superior (CAPES), Fundacao Universitaria Jose Bonifacio (FUJB) and Programa de Nucleos de Excelencia (PRONEX). The financial support through the projects: PTDC/SAUSAP/113196/2009/ FCOMP-01-0124-FEDER-016012; PEst-OE/EQB/LA0023/2013; 'BioHealth-Biotechnology and Bioengineering approaches to improve health quality', NORTE-07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte, QREN; RECI/BBB-EBI/0179/2012/FCOMP-01-0124-FEDER-027462

Genetic regulation of pituitary gland development in human and mouse

Normal hypothalamopituitary development is closely related to that of the forebrain and is dependent upon a complex genetic cascade of transcription factors and signaling molecules that may be either intrinsic or extrinsic to the developing Rathke’s pouch. These factors dictate organ commitment, cell differentiation, and cell proliferation within the anterior pituitary. Abnormalities in these processes are associated with congenital hypopituitarism, a spectrum of disorders that includes syndromic disorders such as septo-optic dysplasia, combined pituitary hormone deficiencies, and isolated hormone deficiencies, of which the commonest is GH deficiency. The highly variable clinical phenotypes can now in part be explained due to research performed over the last 20 yr, based mainly on naturally occurring and transgenic animal models. Mutations in genes encoding both signaling molecules and transcription factors have been implicated in the etiology of hypopituitarism, with or without other syndromic features, in mice and humans. To date, mutations in known genes account for a small proportion of cases of hypopituitarism in humans. However, these mutations have led to a greater understanding of the genetic interactions that lead to normal pituitary development. This review attempts to describe the complexity of pituitary development in the rodent, with particular emphasis on those factors that, when mutated, are associated with hypopituitarism in humans

Quantitative image analysis of polyhydroxyalkanoates inclusions from microbial mixed cultures under different SBR operation strategies

Polyhydroxyalkanoates (PHAs) produced from mixed microbial cultures (MMC), regarded as potential substitutes of petrochemical plastics, can be found as intracellular granules in various microorganisms under limited nutrient conditions and excess of carbon source. PHA is traditionally quantified by laborious and time-consuming chromatography analysis, and a simpler and faster method to assess PHA contents from MMC, such as quantitative image analysis (QIA), is of great interest. The main purpose of the present work was to upgrade a previously developed QIA methodology (Mesquita et al., 2013a, 2015) for MMC intracellular PHA contents quantification, increase the studied intracellular PHA concentration range and extend to different sequencing batch reactor (SBR) operation strategies. Therefore, the operation of a new aerobic dynamic feeding (ADF) SBR allowed further extending the studied operating conditions, dataset, and range of the MMC intracellular PHA contents from the previously reported anaerobic/aerobic cycle SBR. Nile Blue A (NBA) staining was employed for epifluorescence microscope visualization and image acquisition, further fed to a custom developed QIA. Data from each of the feast and famine cycles of both SBR were individually processed using chemometrics analysis, obtaining the correspondent partial least squares (PLS) models. The PHA concentrations determined from PLS models were further plotted against the results obtained in the standard chromatographic method. For both SBR the predicted ability was higher at the end of the feast stage than for the famine stage. Indeed, an independent feast and famine QIA data treatment was found to be fundamental to obtain the best prediction abilities. Furthermore, a promising overall correlation (R2 of 0.83) could be found combining the overall QIA data regarding the PHA prediction up to a concentration of 1785.1 mgL-1 (37.3 wt%). Thus, the results confirm that the presented QIA methodology can be seen as promising for estimating higher intracellular PHA concentrations for a larger reactors operation systems and further extending the prediction range of previous studies.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE01-0145-FEDER-000004) funded by European Regional Development Fundunder the scope ofNorte2020 - ProgramaOperacional Regional do Norte.The authors also acknowledge the financial support to Cristiano S. Leal (PTDC/EBB-EBI/103147/2008, FCOMP-01-0124-FEDER009704) and Daniela P. Mesquita through the FCT postdoctoral grant (SFRH/BPD/82558/2011).info:eu-repo/semantics/publishedVersio

Improved Survival, Vascular Differentiation and Wound Healing Potential of Stem Cells Co-Cultured with Endothelial Cells

In this study, we developed a methodology to improve the survival, vascular differentiation and regenerative potential of umbilical cord blood (UCB)-derived hematopoietic stem cells (CD34+ cells), by co-culturing the stem cells in a 3D fibrin gel with CD34+-derived endothelial cells (ECs). ECs differentiated from CD34+ cells appear to have superior angiogenic properties to fully differentiated ECs, such as human umbilical vein endothelial cells (HUVECs). Our results indicate that the pro-survival effect of CD34+-derived ECs on CD34+ cells is mediated, at least in part, by bioactive factors released from ECs. This effect likely involves the secretion of novel cytokines, including interleukin-17 (IL-17) and interleukin-10 (IL-10), and the activation of the ERK 1/2 pathway in CD34+ cells. We also show that the endothelial differentiation of CD34+ cells in co-culture with CD34+-derived ECs is mediated by a combination of soluble and insoluble factors. The regenerative potential of this co-culture system was demonstrated in a chronic wound diabetic animal model. The co-transplantation of CD34+ cells with CD34+-derived ECs improved the wound healing relatively to controls, by decreasing the inflammatory reaction and increasing the neovascularization of the wound

Ezrin enrichment on curved membranes requires a specific conformation or interaction with a curvature-sensitive partner

One challenge in cell biology is to decipher the biophysical mechanisms governing protein enrichment on curved membranes and the resulting membrane deformation. The ERM protein ezrin is abundant and associated with cellular membranes that are flat, positively or negatively curved. Using in vitro and cell biology approaches, we assess mechanisms of ezrin's enrichment on curved membranes. We evidence that wild-type ezrin (ezrinWT) and its phosphomimetic mutant T567D (ezrinTD) do not deform membranes but self-assemble antiparallelly, zipping adjacent membranes. EzrinTD's specific conformation reduces intermolecular interactions, allows binding to actin filaments, which reduces membrane tethering, and promotes ezrin binding to positively-curved membranes. While neither ezrinTD nor ezrinWT senses negative curvature alone, we demonstrate that interacting with curvature-sensing I-BAR-domain proteins facilitates ezrin enrichment in negatively-curved membrane protrusions. Overall, our work demonstrates that ezrin can tether membranes, or be targeted to curved membranes, depending on conformations and interactions with actin and curvature-sensing binding partners.Peer reviewe

Stress response of lettuce (Lactuca sativa) to environmental contamination with selected pharmaceuticals: A proteomic study

Pharmaceutical compounds have been found in rivers and treated wastewaters. They often contaminate irrigation waters and consequently accumulate in edible vegetables, causing changes in plants metabolism The main objective of this work is to understand how lettuce plants cope with the contamination from three selected pharmaceuticals using a label free proteomic analysis. A lettuce hydroponic culture, grown for 36 days, was exposed to metformin, acetaminophen and carbamazepine (at 1 mg/L), during 8 days, after which roots and leaves were sampled and analysed using a liquid chromatography-mass spectrometry proteomics-based approach. In roots, a total of 612 proteins showed differentially accumulation while in leaves 237 proteins were identified with significant differences over controls. Carbamazepine was the contaminant that most affected protein abundance in roots, while in leaves the highest number of differentially accumulated proteins was observed for acetaminophen. In roots under carbamazepine, stress related protein species such as catalase, superoxide dismutase and peroxidases presented higher abundance. Ascorbate peroxidase increased in roots under metformin. Cell respiration protein species were affected by the presence of the three pharmaceuticals suggesting possible dysregulation of the Krebs cycle. Acetaminophen caused the main differences in respiration pathways, with more emphasis in leaves. Lettuce plants revealed different tolerance levels when contaminants were compared, being more tolerant to metformin presence and less tolerant to carbamazepineinfo:eu-repo/semantics/acceptedVersio