Global mRNA expression analysis in myosin II deficient strains of Saccharomyces cerevisiae reveals an impairment of cell integrity functions

<p>Abstract</p> <p>Background</p> <p>The <it>Saccharomyces cerevisiae MYO1 </it>gene encodes the myosin II heavy chain (Myo1p), a protein required for normal cytokinesis in budding yeast. Myo1p deficiency in yeast (<it>myo1Δ</it>) causes a cell separation defect characterized by the formation of attached cells, yet it also causes abnormal budding patterns, formation of enlarged and elongated cells, increased osmotic sensitivity, delocalized chitin deposition, increased chitin synthesis, and hypersensitivity to the chitin synthase III inhibitor Nikkomycin Z. To determine how differential expression of genes is related to these diverse cell wall phenotypes, we analyzed the global mRNA expression profile of <it>myo1Δ </it>strains.</p> <p>Results</p> <p>Global mRNA expression profiles of <it>myo1Δ </it>strains and their corresponding wild type controls were obtained by hybridization to yeast oligonucleotide microarrays. Results for selected genes were confirmed by real time RT-PCR. A total of 547 differentially expressed genes (p ≤ 0.01) were identified with 263 up regulated and 284 down regulated genes in the <it>myo1Δ </it>strains. Gene set enrichment analysis revealed the significant over-representation of genes in the protein biosynthesis and stress response categories. The <it>SLT2/MPK1 </it>gene was up regulated in the microarray, and a <it>myo1Δslt2Δ </it>double mutant was non-viable. Overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31 </it>suppressed the hypersensitivity to Nikkomycin Z and increased the levels of phosphorylated Slt2p in <it>myo1Δ </it>strains. Increased levels of phosphorylated Slt2p were also observed in wild type strains under these conditions.</p> <p>Conclusion</p> <p>Following this analysis of global mRNA expression in yeast <it>myo1Δ </it>strains, we conclude that 547 genes were differentially regulated in <it>myo1Δ </it>strains and that the stress response and protein biosynthesis gene categories were coordinately regulated in this mutant. The <it>SLT2/MPK1 </it>gene was confirmed to be essential for <it>myo1Δ </it>strain viability, supporting that the up regulated stress response genes are regulated by the <it>PKC1 </it>cell integrity pathway. Suppression of Nikkomycin Z hypersensitivity together with Slt2p phosphorylation was caused by the overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31</it>. These ribosomal protein mRNAs were down regulated in the <it>myo1Δ </it>arrays, suggesting that down regulation of ribosomal biogenesis may affect cell integrity in <it>myo1Δ </it>strains.</p

The neuropilin 1 cytoplasmic domain is required for VEGF-A-dependent arteriogenesis.

Neuropilin 1 (NRP1) plays an important but ill-defined role in VEGF-A signaling and vascular morphogenesis. We show that mice with a knockin mutation that ablates the NRP1 cytoplasmic tail (Nrp1(cyto)) have normal angiogenesis but impaired developmental and adult arteriogenesis. The arteriogenic defect was traced to the absence of a PDZ-dependent interaction between NRP1 and VEGF receptor 2 (VEGFR2) complex and synectin, which delayed trafficking of endocytosed VEGFR2 from Rab5+ to EAA1+ endosomes. This led to increased PTPN1 (PTP1b)-mediated dephosphorylation of VEGFR2 at Y(1175), the site involved in activating ERK signaling. The Nrp1(cyto) mutation also impaired endothelial tubulogenesis in vitro, which could be rescued by expressing full-length NRP1 or constitutively active ERK. These results demonstrate that the NRP1 cytoplasmic domain promotes VEGFR2 trafficking in a PDZ-dependent manner to regulate arteriogenic ERK signaling and establish a role for NRP1 in VEGF-A signaling during vascular morphogenesis

Plants Probiotics as a Tool to Produce Highly Functional Fruits: The Case of Phyllobacterium and Vitamin C in Strawberries

10 páginas, 1 tabla, 1 figuraThe increasing interest in the preservation of the environment and the health of consumers is changing production methods and food consumption habits. Functional foods are increasingly demanded by consumers because they contain bioactive compounds involved in health protection. In this sense biofertilization using plant probiotics is a reliable alternative to the use of chemical fertilizers, but there are few studies about the effects of plant probiotics on the yield of functional fruits and, especially, on the content of bioactive compounds. In the present work we reported that a strain of genus Phyllobacterium able to produce biofilms and to colonize strawberry roots is able to increase the yield and quality of strawberry plants. In addition, the fruits from plants inoculated with this strain have significantly higher content in vitamin C, one of the most interesting bioactive compounds in strawberries. Therefore the use of selected plant probiotics benefits the environment and human health without agronomical losses, allowing the production of highly functional foods.This work was granted by “Junta de Castilla y León” (Regional Government, Grant SA183A11-2) and MINECO (Central Government, Grant AGL2011-29227). Paula García-Fraile is recipient of a postdoctoral researcher contract from Academy of Sciences of the Czech Republic. José David Flores-Félix was supported by a fellowship of Salamanca University. Marta Marcos García was supported by a fellowship of Fundación Miguel Casado San José. Luís R. Silva is grateful to the financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project Pest-C/EQB/LA0006/2013 and from the European Union (FEDER funds) under the framework of QREN through Project NORTE-07-0124-FEDER- 000069.Peer reviewe

Biodiversidad de parásitos Micropterus salmoides en la presa La Boca (Rodrigo Gómez), en el estado de Nuevo león, México.

Los estudios referentes al conocimiento de la helmintofauna de Micropterus salmoides son recientes en el estado de Nuevo León, México. En el presente estudio se identificaron a los metazoarios parásitos de la lobina negra. Se calculó la prevalencia, intensidad y abundancia y su relación estadística con el peso, la talla y el sexo en la presa La Boca. Se realizaron colectas mensuales de enero del 2011 a enero de 2012. Se capturaron 116 peces en total, mediante diferentes artes de pesca. Todos los peces fueron positivos para al menos una especie de helminto, identificándose un total de cinco grupos de metazoarios parásitos: monogeneos con dos géneros (Cleidodiscus floridanus y Acolpenteron ureteroecetes; trematodo digeneo (Neascus vancleavei), un céstodo (Proteocephalus ambloplitis); un nematodo (Contracaecum sp.), un acantocéfalo (Neoechinorhynchus cylindratus); un copépodo (Ergasilus versicolor). Las especies más importantes por los valores de abundancia y prevalencia fueron los tremátodos Neascus vancleavei y Cleidodiscus floridanus y las larvas del nematodo Contracaecum sp

Frequency of CYP2C9 (*2, *3 and IVS8‑109A>T) allelic variants, and their clinical implications, among Mexican patients with diabetes mellitus type 2 undergoing treatment with glibenclamide and metformin

The majority of Mexican patients with diabetes mellitus type 2 (DMT2) (67.9-85.0%) are prescribed sulpho nylureas (SUs), which are metabolized by cytochrome P450 2C9 (abbreviated as CYP2C9). SUs are a type of oral anti-diabetic compound which inhibit ATP-sensitive potas sium channels, thus inducing glucose-independent insulin release by the β-pancreatic cells. The wide variability reported in SU responses has been attributed to the polymorphisms of CYP2C9. The present study aimed to describe CYP2C9 polymorphisms (*2, *3 and IVS8‑109T) within a sample of Mexican patients with DMT2, while suggesting the potential clinical implications in terms of glibenclamide response vari ability. From a sample of 248 patients with DMT2 who initially consented to be studied, those ultimately included in the study were treated with glibenclamide (n=11), glibenclamide combined with metformin (n=112) or metformin (n=76), and were subsequently genotyped using a reverse transcrip tion-quantitative polymerase chain reaction (PCR), end-point allelic discrimination and PCR amplifying enzymatic restric tion fragment long polymorphism. Clinical data were gathered through medical record revision. The frequencies revealed were as follows: CYP2C9*1/*1, 87.5%; *1/*2, 6.5%; *1/*3, 5.2%; and CYP2C9, IVS8‑109A>T, 16.1%. Glibenclamide significantly reduced the level of pre‑prandial glucose (P<0.01) and the percentage of glycated hemoglobin (%HbA1c; P<0.01) for IVS8‑109A>T compared with combined glibenclamide and metformin treatment. Concerning the various treatments with respect to the different genotypes, the percentages obtained were as follows: Glibenclamide A/A, HbA1c<6.5=33.3%; glibenclamide + metformin A/A, HbA1c<6.5=24.6%; gliben clamide A/T, HbA1c<6.5=33.3%; glibenclamide + metformin A/T, HbA1c<6.5=25%; glibenclamide T/T, HbA1c<6.5=100%; and glibenclamide + metformin T/T, HbA1c<6.5=12.5%. Altogether, these results revealed that, although genetically customized prescriptions remain a desirable goal to increase the chances of therapeutic success, within the studied popu lation neither allelic variants nor dosages demonstrated a clear association with biomarker levels. A key limitation of the present study was the lack of ability to quantify either the plasma concentrations of SU or their metabolites; therefore, further, precise experimental and observational studies are required

Level of blood pressure control in a hypertensive population when measurements are performed outside the clinical setting

Background: To determine whether the number of optimally controlled hypertensive patients is higher using self-measurement of blood pressure at home and ambulatory monitoring, compared to using conventional blood pressure measurements at the doctor&#8217;s office. Method: An observational, cross-sectional, multicentre, descriptive study of a random sample of 237 primary health care patients, known to be hypertensive, from Badajoz (Spain). Blood pressure was measured at the doctor&#8217;s office and by self-measurement at home. Those patients showing good control by self-measurement were subjected to 24-hour ambulatory monitoring. Optimal control was understood as blood pressure < 140/90 mm Hg when measured at the doctor&#8217;s office, and < 135/85 mm Hg when self-measured at home and by daytime ambulatory monitoring. Results: Mean systolic/diastolic measurements at the doctor&#8217;s office and by self-measurement were 145.6/83.9 and 134.0/78.7 mm Hg, respectively (p < 0.000). In the population optimally controlled by self-measurement and who subsequently received ambulatory monitoring, the mean blood pressure was 121.8/73.4 and 125.6/76.2 mm Hg, respectively (p = 0.002; p < 0.000). When measured at the doctor&#8217;s office blood pressure was controlled in about 29.5% (95% CI 23.7-35.3%) of patients, in 38% when self-measured (95% CI 31.4-44.2%; p < 0.000), and in 24.5% when it was confirmed through ambulatory monitoring (95% CI 15.4-33.6%). Sensitivity and positive predictive values of the office measurements for the detection of patients who were well-controlled by self-measurement were 50% and 64.3%, respectively, and 53.4% and 73.8% as regards ambulatory monitoring. Conclusions: A higher level of control is achieved with self-measurement at home not confirmed by ambulatory monitoring. Therefore, the white coat effect does not seem to influence the percentage of well-controlled patients detected at the doctor&#8217;s office. Office blood pressure does not appear to be useful in distinguishing which individual patients are optimally controlled

Pharmacogenetics of Metformin Transporters Suggests No Association with Therapeutic Inefficacy among Diabetes Type 2 Mexican Patients

Mexico has been under official epidemiological alert due to diabetes since 2016. This study presents new information on the frequency and variants of metformin transporters OCT1, OCT2, OCT3, ABCB1, and CYP2C9 variants as well. It also reports the association with HbA1c control on 103 DMT2 patients. They were genotyped through real-time PCR (TaqMan assays) and grouped according to treatment: metformin and metformin + glibenclamide. Metformin plasmatic levels were determined through mass spectrometry. The analysis of HbA1c showed statistical significance across genotypes in polymorphisms rs72552763 (p = 0.022), rs622342 (p = 0.009), rs1128503 (p = 0.021), and rs2032582 (p = 0.009) within the monotherapy group. Bivariate analysis found no association between any polymorphism and HbA1c control. Two logistic regression models accounted for two diplotypes in OCT1 and ABCB1, including statistically significant covariates. The first model yielded significance in age (p = 0.026), treatment period [p = 0.001], BMI ≥ 25 kg/m2 (p = 0.043), and combined therapy (p < 0.001). There was no association with GAT/GAT of rs72552763 or A/A rs622342 in OCT1. The second model yielded significance in age (p = 0.017), treatment period (p = 0.001), BMI ≥ 25 kg/m2 (p = 0.042), and combined therapy (p < 0.001), finding no association with C/C of rs1128503 or G/G of rs2032582 in ABCB1. Our multinomial logistic regression results may benefit future predictive analyses in diabetic populations

Pesquisa de validação da percepção Exame Nacional de Enfermagem do Chile

Investigación descriptiva cuantitativa y cualitativa. Objetivo: validar encuesta de percepción de los egresados de Enfermería en Chile, respecto al Examen Nacional de Enfermería. Metodología: lo cualitativo consideró el método Delphi participando expertos en la disciplina de enfermería y del área educación y evaluación. Para lo cuantitativo se utilizó el análisis de alfa de Cronbach, usando el sistema SPSS. Resultados: El K obtenido por los expertos estuvo entre el 0,95 a 1 lo que determinó que todos tenían un alto grado de competencia en el tema sobre el cual se les consultó. Los expertos vincularon los objetivos propuestos con cada uno de los 21 ítems de la encuesta. En 8 de ellos el 100% de los expertos coincidió con lo plateado por la Comisión del Examen Nacional de Enfermería, en 10 de los ítems un 80%, en 2 de los ítems un 50% y en 1 ítem un 33%. El alpha de Cronbach obtenido fue de 0,97, lo que demuestra un alto grado de consistencia interna del instrumento. Conclusiones: Se valida la encuesta de opinión, cumpliendo de manera rigurosa y lógica con todas sus etapas, demostrando su validez y confiabilidad.Quantitative and qualitative descriptive research. Objective: to validate the survey on the perception of nursing graduates in Chile, regarding the National Exam taken and applied. Methodology: qualitative, using the Delphi method with experts in the areas of nursing, education, and evaluation; and quantitative, using Cronbach's alpha analysis with the SPSS system. Results: The K was between 0.95 to 1, which suggests that all of the experts had a high degree of competence in the opinion topic. The objectives proposed in the 21-item survey were linked. In 8 items 100% of the experts stated their agreement with the aspects raised by the National Nursing Exam Commission; in 10 items 80%, in 2 items 50% and in 1 item 33%. Cronbach's alpha method estimated the internal consistency reliability of the instrument to be 0.97, demonstrating a high degree of internal consistency among the items analyzed. Conclusions: to validate an instrument it is necessary to rigorously complete each step with a sequence that allows one to account for validity and reliability

Trazodone for the treatment of fibromyalgia: an open-label, 12-week study

Background: Despite its frequent use as a hypnotic, trazodone has not been systematically assessed in fibromyalgia patients. In the present study have we evaluated the potential effectiveness and tolerability of trazodone in the treatment of fibromyalgia. Methods: A flexible dose of trazodone (50-300 mg/day), was administered to 66 fibromyalgia patients for 12 weeks. The primary outcome measure was the Pittsburgh Sleep Quality Index (PSQI). Secondary outcome measures included the Fibromyalgia Impact Questionnaire (FIQ), the Beck Depression Inventory (BDI), the Hospital Anxiety and Depression Scale (HADS), the Brief Pain Inventory (BPI), the Short-Form Health Survey (SF-36), and the Patients' Global Improvement Scale (PGI). Trazodone's emergent adverse reactions were recorded. Data were analyzed with repeated measures one-way ANOVA and paired Student's t test. Results: Trazodone markedly improved sleep quality, with large effect sizes in total PSQI score as well on sleep quality, sleep duration and sleep efficiency. Significant improvement, although with moderate effect sizes, were also observed in total FIQ scores, anxiety and depression scores (both HADS and BDI), and pain interference with daily activities. Unexpectedly, the most frequent and severe side effect associated with trazodone in our sample was tachycardia, which was reported by 14 (21.2%) patients. Conclusions: In doses higher than those usually prescribed as hypnotic, the utility of trazodone in fibromyalgia management surpasses its hypnotic activity. However, the emergence of tachycardia should be closely monitored. Trial registration: This trial has been registered with ClinicalTrials.gov number NCT-00791739