Kinase Inhibitor Profile For Human Nek1, Nek6, And Nek7 And Analysis Of The Structural Basis For Inhibitor Specificity.

Human Neks are a conserved protein kinase family related to cell cycle progression and cell division and are considered potential drug targets for the treatment of cancer and other pathologies. We screened the activation loop mutant kinases hNek1 and hNek2, wild-type hNek7, and five hNek6 variants in different activation/phosphorylation statesand compared them against 85 compounds using thermal shift denaturation. We identified three compounds with significant Tm shifts: JNK Inhibitor II for hNek1(Δ262-1258)-(T162A), Isogranulatimide for hNek6(S206A), andGSK-3 Inhibitor XIII for hNek7wt. Each one of these compounds was also validated by reducing the kinases activity by at least 25%. The binding sites for these compounds were identified by in silico docking at the ATP-binding site of the respective hNeks. Potential inhibitors were first screened by thermal shift assays, had their efficiency tested by a kinase assay, and were finally analyzed by molecular docking. Our findings corroborate the idea of ATP-competitive inhibition for hNek1 and hNek6 and suggest a novel non-competitive inhibition for hNek7 in regard to GSK-3 Inhibitor XIII. Our results demonstrate that our approach is useful for finding promising general and specific hNekscandidate inhibitors, which may also function as scaffolds to design more potent and selective inhibitors.

Pharmacological Characterization Of The Relaxation Induced By The Soluble Guanylate Cyclase Activator, Bay 60-2770 In Rabbit Corpus Cavernosum.

To characterize the relaxation induced by the soluble guanylate cyclase (sGC) activator, BAY 60-2770 in rabbit corpus cavernosum. Penis from male New Zealand rabbits were removed and fours strips of corpus cavernosum (CC) were obtained. Concentration-response curves to BAY 60-2770 were carried out in the absence and presence of inhibitors of nitric oxide synthase, L-NAME (100 μM), sGC, ODQ (10 μM) and phosphodiestarase type 5, tadalafil (0.1 μM). The potency (pEC50) and maximal response (Emax) values were determined. Second, electrical-field stimulation (EFS)-induced contraction or relaxation was realized in the absence and presence of BAY 60-2770 (0.1 or 1 μM) alone or in combination of ODQ (10 μM). In the case of EFS-induced relaxation two protocols were realized: 1) ODQ (10 μM) was first incubated for 20 min and then BAY 60-2770 (1 μM) was added for another 20 min (ODQ + BAY 60-2770). In different CC strips, BAY 60-2770 was incubated for 20 min followed by another 20 min with ODQ (BAY 60-2770 + ODQ). The intracellular levels of cyclic guanosine monophosphate (cGMP) were also determined. BAY 60-2770 potently relaxed rabbit CC with pEC50 and Emax values of 7.58 ± 0.19 and 81 ± 4%, respectively. The inhibitors ODQ (n=7) or tadalafil (n=7) produced 4.2- and 6.3-leftward shifts, respectively in BAY 60-2770-induced relaxation without interfering on the Emax values. The intracellular levels of cGMP were augmented after stimulation with BAY 60-2770 (1 μM) alone, whereas its co-incubation with ODQ produced even higher levels of cGMP. The EFS-induced contraction was reduced in the presence of BAY 60-2770 (1 μM) and this inhibition was even greater when BAY 60-2770 was co-incubated with ODQ. The nitrergic stimulation induced CC relaxation, which was abolished in the presence of ODQ. BAY 60-2770 alone increased the amplitude of relaxation. Co-incubation of ODQ and BAY 60-2770 did not alter the relaxation in comparison with ODQ alone. Interestingly, when BAY 60-2770 was incubated prior to ODQ, EFS-induced relaxation was partly restored in comparison with ODQ alone or ODQ + BAY 60-2770. Considering that the relaxation induced by the sGC activator, BAY 60-2770 was increased after sGC oxidation and unaltered in the absence of nitric oxide, these class of substances are advantageous over sGC stimulators or PDE5 inhibitors for the treatment in those patients with erectile dysfunction and high endothelial damage. This article is protected by copyright. All rights reserved.

Analise comparativa das caracteristicas cinematicas da coluna durante a marcha e a corrida

Universidade Estadual de Campinas . Faculdade de Educação Física

Estrategias organizacionais para a promoção de saude e qualidade de vida : avaliando a qualidade de vida no trabalho

Universidade Estadual de Campinas . Faculdade de Educação Física

Teste "W" 20 metros : proposta computadorizada para analise da performance especifica em atletas de voleibol

Universidade Estadual de Campinas . Faculdade de Educação Física

Sobre uma degenerescência acidental nos deslocamentos químicos de RMN de 31P{¹H} em complexos difosfínicos de rutênio

The [RuCl(bipy)(dppb)(4-pic)]PF6 complex was prepared and fully characterized. The X-ray crystal structure of this complex was determined in order to make an unambiguous distinction between the two possible positions of the 4-methylpyridine ligand (4-pic) in the compound: trans to phosphorus atom or trans to nitrogen atom. The [RuCl(bipy)(dppb)(4-pic)]PF6 complex exhibits an unusual temperature-dependent accidental degeneracy of the 31P chemical shifts in its solution NMR spectrum.

Effect of temperature on the electro-oxidation of ethanol on platinum

We present in this work an experimental investigation of the effect of temperature (from 25 to 180 ºC) in the electro-oxidation of ethanol on platinum in two different phosphoric acid concentrations. We observed that the onset potential for ethanol electro-oxidation shifts to lower values and the reaction rates increase as temperature is increased for both electrolytes. The results were rationalized in terms of the effect of temperature on the adsorption of reaction intermediates, poisons, and anions. The formation of oxygenated species at high potentials, mainly in the more diluted electrolyte, also contributes to increase the electro-oxidation reaction rate.

Cloning, purification and comparative characterization of two digestive lysozymes from Musca domestica larvae

cDNA coding for two digestive lysozymes (MdL1 and MdL2) of the Musca domestica housefly was cloned and sequenced. MdL2 is a novel minor lysozyme, whereas MdL1 is the major lysozyme thus far purified from M. domestica midgut. MdL1 and MdL2 were expressed as recombinant proteins in Pichia pastoris, purified and characterized. The lytic activities of MdL1 and MdL2 upon Micrococcus lysodeikticus have an acidic pH optimum (4.8) at low ionic strength (μ = 0.02), which shifts towards an even more acidic value, pH 3.8, at a high ionic strength (μ = 0.2). However, the pH optimum of their activities upon 4-methylumbelliferyl N-acetylchitotrioside (4.9) is not affected by ionic strength. These results suggest that the acidic pH optimum is an intrinsic property of MdL1 and MdL2, whereas pH optimum shifts are an effect of the ionic strength on the negatively charged bacterial wall. MdL2 affinity for bacterial cell wall is lower than that of MdL1. Differences in isoelectric point (pI) indicate that MdL2 (pI = 6.7) is less positively charged than MdL1 (pI = 7.7) at their pH optima, which suggests that electrostatic interactions might be involved in substrate binding. In agreement with that finding, MdL1 and MdL2 affinities for bacterial cell wall decrease as ionic strength increases.

Associação entre prática de atividade física, escolaridade e perfil alimentar de motoristas de caminhão

A demanda da sociedade moderna intensificou o trabalho ininterrupto em diversas profissões. Além disso, o modo de organização da sociedade, com atividades predominantemente mecanizadas, tem contribuído para a prevalência de hábitos de vida não saudáveis, como a inatividade física. Este estudo objetiva verificar se fatores ocupacionais, sociodemográficos, antropométricos e alimentares estão associados à prática de atividade física insuficiente e se há diferenças nessa associação entre motoristas de caminhão que trabalham de dia ou à noite. Participaram da pesquisa 470 motoristas de caminhão, que responderam a questionários de dados sociodemográficos, atividade física e frequência alimentar. Foi realizada uma regressão logística univariada para verificar a associação entre atividade física insuficiente e as demais variáveis. Além disso, a regressão logística múltipla foi testada para obter modelos que mostrem a associação de conjuntos de variáveis relacionados à atividade física insuficiente. Os resultados indicaram que a prática de atividade física está associada ao maior nível de escolaridade (OR = 1,84; IC = 1,22-2,76) e menor consumo de bebidas alcoólicas (OR = 1,59; IC = 1,04-2,45). Maior ingestão de cereais integrais (OR = 1,63; IC = 1,08-2,46) foi associada à prática regular de atividade física. Entre os trabalhadores noturnos, foi encontrada associação entre a prática regular de atividade física, maior consumo de cereais integrais (OR = 2,02; IC =1,13-3,60) e menor consumo de carboidratos simples (OR = 1,91; IC = 1,08-3,37).

Influence of the bipartite scrotum on the testicular and scrotal temperatures in goats

The influence of the scrotal bipartition and of the year period on the scrotal-testicular thermal regulation was evaluated in male goats raised in Piaui State, Brazil. Eighteen male goats at mating age were accomplished in this study and arranged into three Groups (6 animals each) obeying the classification as goats presenting no scrotal bipartition (Group I), goats showing scrotal bipartition at 50% of the testicular length (Group II), and goats with more than 50% of scrotal bipartition (Group III). The scrotal, testicular and spermatic funiculi temperatures were evaluated invasively with the aid of a digital thermometer and non-invasive with a pyrometer in the proximal, medial and distal portion. The data were acquired during the dry (October-November) and rainy (February-March) period of the year, measured in two shifts: morning (6h00-7h00) and afternoon (14h00-15h00). The results were submitted to variance analysis (ANOVA) following the SNK test for average comparison (p<0.05). The year period interfered on the scrotal-testicular thermal regulation, due to increased temperatures of the scrotal, testicular and spermatic funiculi during the dry period in comparison with the rainy period. The bipartition level was also a factor which contributed to the influence of scrotal-testicular temperature regulation, due to lower average scrotal-testicular temperature rates observed during both periods in the goats with higher levels of scrotal bipartition (>50%). It is possible to conclude that with the experimental conditions applied on this study, the level of scrotal bipartition and the climatic conditions interfere with the scrotal-testicular thermal regulation in goats.

Qualidade de vida associada a saúde e condições de trabalho entre profissionais de enfermagem

OBJETIVO: Avaliar condições de trabalho associadas à qualidade de vida relacionada à saúde entre profissionais de enfermagem. MÉTODOS: Estudo transversal realizado em um hospital universitário de São Paulo, SP, em 2004-2005. A população estudada foi de 696 enfermeiros, técnicos e auxiliares de enfermagem, predominantemente feminina (87,8 por cento) e que trabalhava em turnos diurnos e/ou noturnos. Os dados sociodemográficos, de condições de trabalho e de vida, hábitos de vida e sintomas de saúde auto-referidos foram obtidos por meio de questionários auto-aplicados: Resultados de Estudos de Saúde - versão reduzida, Escala de Estresse no Trabalho e Desequilíbrio Esforço-Recompensa. Valores do coeficiente 1,01 significam mais esforços do que recompensas no trabalho. Modelos de regressão logística ordinal de chances proporcionais foram ajustados para cada dimensão do SF-36. RESULTADOS: Aproximadamente 22 por cento da população foi classificada como trabalhando em condições de alto desgaste e 8 por cento com mais esforços do que recompensas no trabalho. As dimensões com piores escores médios no SF-36 foram vitalidade, dor e saúde mental. Alto desgaste no trabalho, ter mais esforços que recompensas e ser enfermeira associaram-se de maneira independente aos baixos escores da dimensão de aspectos emocionais. As dimensões relacionadas à saúde mental foram as que mais sofreram influência dos fatores psicossociais do trabalho. CONCLUSÕES: Apresentar mais esforços do que recompensas no trabalho foi mais significativo para a qualidade de vida associada à saúde do que o alto desgaste no trabalho (altas demandas e baixo controle). Os resultados indicam que a análise conjunta dos fatores psicossociais de desequilíbrio esforço-recompensa e demanda-controle contribuiu para a discussão sobre os papéis profissionais, condições de trabalho e qualidade de vida relacionada à saúde de profissionais de enfermagem

Work and excessive sleepiness among Brazilian evening high school students: effects on days off

Previous studies have revealed that students who work and study build up sleep deficits during the wrkweek, which can trigger a sleep rebound during days off. The objective of this study was to investigate the impact of working on sleepiness during days off working / non-working on sleepiness days off among high school students. The study population, aged 14-21 years, attended evening classes in São Paulo, Brazil. For the study, the students completed questionaires on living conditions, health, and work; wore actigraphs; and completed the Karolinska Sleepiness Scale (KSS). To predict sleepiness, a logistic regression analysis was performed. Excessive sleepiness was observed on the first day off among working students. Results suggest that working is a significant predictor for sleepiness and that two shifts of daily systematic activities, study and work, might lead to excessive daytime sleepiness on the first day off. Further, this observed excessive sleepiness may reflect the sleep debt accumulated during the workweek

Effects of naps at work on the sleepiness of 12-hour night shift nursing personnel

Background and objective: The purpose of the present study was to evaluate the effects of a nap at work on the sleepiness of 12-hour, night-shift (registered and assistant) nursing personnel.Methods: Twelve nurses filled out daily logs, the Karolinska Sleepiness Scale (KS), and wore wrist actigraphs for two periods of four continuous days.Results: Mean nap duration during the night shifts was 138.3 (SD+39.8) minutes. The mean sleepiness level assessed by the KS score was lower, 3.3 (SD±1.6), when the nap was taken during the first span (00:01 - 03:00h) of the night shift, compared with 6.6 (SD±1.0) when there was no nap. The mean sleepiness level assessed by the KS score was also lower, 3.6 (SD±0.9), when the nap was taken during the second span (03:01 - 06:00h) of the night shift, compared with 7.0 (SD±1.1) when there was no nap. Thus, napping either during the first or second part of the night shift reduces sleepiness of 12-hour, night-shift nursing personnel. Moreover, the mean duration of the first sleep episode after night work was longer in those who did not nap than in those who did. Conclusions: The results of this study show that napping during the 12-hour, night-shift results in less sleepiness at work and less need for recovery sleep after work

Precision of distances and ordering of microsatellite markers in consensus linkage maps of chromosomes 1, 3 and 4 from two reciprocal chicken populations using bootstrap sampling

Some factors complicate comparisons between linkage maps from different studies. This problem can be resolved if measures of precision, such as confidence intervals and frequency distributions, are associated with markers. We examined the precision of distances and ordering of microsatellite markers in the consensus linkage maps of chromosomes 1, 3 and 4 from two F 2 reciprocal Brazilian chicken populations, using bootstrap sampling. Single and consensus maps were constructed. The consensus map was compared with the International Consensus Linkage Map and with the whole genome sequence. Some loci showed segregation distortion and missing data, but this did not affect the analyses negatively. Several inversions and position shifts were detected, based on 95% confidence intervals and frequency distributions of loci. Some discrepancies in distances between loci and in ordering were due to chance, whereas others could be attributed to other effects, including reciprocal crosses, sampling error of the founder animals from the two populations, F(2) population structure, number of and distance between microsatellite markers, number of informative meioses, loci segregation patterns, and sex. In the Brazilian consensus GGA1, locus LEI1038 was in a position closer to the true genome sequence than in the International Consensus Map, whereas for GGA3 and GGA4, no such differences were found. Extending these analyses to the remaining chromosomes should facilitate comparisons and the integration of several available genetic maps, allowing meta-analyses for map construction and quantitative trait loci (QTL) mapping. The precision of the estimates of QTL positions and their effects would be increased with such information.

Transcriptional profiling reveals the expression of novel genes in response to various stimuli in the human dermatophyte Trichophyton rubrum

Background: Cutaneous mycoses are common human infections among healthy and immunocompromised hosts, and the anthropophilic fungus Trichophyton rubrum is the most prevalent microorganism isolated from such clinical cases worldwide. The aim of this study was to determine the transcriptional profile of T. rubrum exposed to various stimuli in order to obtain insights into the responses of this pathogen to different environmental challenges. Therefore, we generated an expressed sequence tag (EST) collection by constructing one cDNA library and nine suppression subtractive hybridization libraries. Results: The 1388 unigenes identified in this study were functionally classified based on the Munich Information Center for Protein Sequences (MIPS) categories. The identified proteins were involved in transcriptional regulation, cellular defense and stress, protein degradation, signaling, transport, and secretion, among other functions. Analysis of these unigenes revealed 575 T. rubrum sequences that had not been previously deposited in public databases. Conclusion: In this study, we identified novel T. rubrum genes that will be useful for ORF prediction in genome sequencing and facilitating functional genome analysis. Annotation of these expressed genes revealed metabolic adaptations of T. rubrum to carbon sources, ambient pH shifts, and various antifungal drugs used in medical practice. Furthermore, challenging T. rubrum with cytotoxic drugs and ambient pH shifts extended our understanding of the molecular events possibly involved in the infectious process and resistance to antifungal drugs.