Bacterial variations on the methionine salvage pathway.

BACKGROUND: The thiomethyl group of S-adenosylmethionine is often recycled as methionine from methylthioadenosine. The corresponding pathway has been unravelled in Bacillus subtilis. However methylthioadenosine is subjected to alternative degradative pathways depending on the organism. RESULTS: This work uses genome in silico analysis to propose methionine salvage pathways for Klebsiella pneumoniae, Leptospira interrogans, Thermoanaerobacter tengcongensis and Xylella fastidiosa. Experiments performed with mutants of B. subtilis and Pseudomonas aeruginosa substantiate the hypotheses proposed. The enzymes that catalyze the reactions are recruited from a variety of origins. The first, ubiquitous, enzyme of the pathway, MtnA (methylthioribose-1-phosphate isomerase), belongs to a family of proteins related to eukaryotic intiation factor 2B alpha. mtnB codes for a methylthioribulose-1-phosphate dehydratase. Two reactions follow, that of an enolase and that of a phosphatase. While in B. subtilis this is performed by two distinct polypeptides, in the other organisms analyzed here an enolase-phosphatase yields 1,2-dihydroxy-3-keto-5-methylthiopentene. In the presence of dioxygen an aci-reductone dioxygenase yields the immediate precursor of methionine, ketomethylthiobutyrate. Under some conditions this enzyme produces carbon monoxide in B. subtilis, suggesting a route for a new gaseous mediator in bacteria. Ketomethylthiobutyrate is finally transaminated by an aminotransferase that exists usually as a broad specificity enzyme (often able to transaminate aromatic aminoacid keto-acid precursors or histidinol-phosphate). CONCLUSION: A functional methionine salvage pathway was experimentally demonstrated, for the first time, in P. aeruginosa. Apparently, methionine salvage pathways are frequent in Bacteria (and in Eukarya), with recruitment of different polypeptides to perform the needed reactions (an ancestor of a translation initiation factor and RuBisCO, as an enolase, in some Firmicutes). Many are highly dependent on the presence of oxygen, suggesting that the ecological niche may play an important role for the existence and/or metabolic steps of the pathway, even in phylogenetically related bacteria. Further work is needed to uncover the corresponding steps when dioxygen is scarce or absent (this is important to explore the presence of the pathway in Archaea). The thermophile T. tengcongensis, that thrives in the absence of oxygen, appears to possess the pathway. It will be an interesting link to uncover the missing reactions in anaerobic environments.

Genome-wide prediction of matrix attachment regions that increase gene expression in mammalian cells.

Gene transfer in eukaryotic cells and organisms suffers from epigenetic effects that result in low or unstable transgene expression and high clonal variability. Use of epigenetic regulators such as matrix attachment regions (MARs) is a promising approach to alleviate such unwanted effects. Dissection of a known MAR allowed the identification of sequence motifs that mediate elevated transgene expression. Bioinformatics analysis implied that these motifs adopt a curved DNA structure that positions nucleosomes and binds specific transcription factors. From these observations, we computed putative MARs from the human genome. Cloning of several predicted MARs indicated that they are much more potent than the previously known element, boosting the expression of recombinant proteins from cultured cells as well as mediating high and sustained expression in mice. Thus we computationally identified potent epigenetic regulators, opening new strategies toward high and stable transgene expression for research, therapeutic production or gene-based therapies.

Nutrição mineral de populações e híbridos de coqueiro (Cocos nucifera L.) cultivados em Bebedouro (SP)

Avaliou-se o estado nutricional de sete genótipos de coqueiro (cinco populações: Anão Amarelo de Gramame, Anão Amarelo da Malásia, Anão Vermelho de Gramame, Anão Verde do Jiqui e Gigante Brasileiro da Praia do Forte e dois híbridos: Anão Amarelo de Gramame x Gigante do Oeste Africano e Anão Vermelho de Gramame x Gigante Brasileiro da Praia do Forte), com 2 anos e 4 meses de idade, plantados em Bebedouro (SP). Com o objetivo de monitorar o estado nutricional das plantas e determinar os efeitos do cultivo sobre a fertilidade do solo, analisaram-se tecido vegetal de folhas com idades diferentes (folhas 4 e 9) e solo. Amostras de folha 4, em comparação com as de folha 9, foram mais eficientes para discriminar as populações e híbridos de coqueiro quanto ao seu estado nutricional. As maiores limitações nutricionais dos genótipos em estudo foram as de zinco, potássio, nitrogênio e cobre. Aparentemente, todas as populações e híbridos observados estavam adequadamente nutridos com fósforo, cálcio, boro, ferro e manganês. Como medida mais urgente para a melhoria do estado nutricional das plantas, recomenda-se aumentar a aplicação de Zn. Os principais efeitos do cultivo de coqueiro sobre a fertilidade relacionaram-se com a acidificação do solo na área de aplicação de adubos.

Efeito da omissão de macronutrientes e boro no crescimento, nos sintomas de deficiências nutricionais e na composição mineral de plantas de camucamuzeiro

Com o objetivo de avaliar o efeito da omissão de macronutrientes e do micronutriente boro no crescimento, nos sintomas de deficiências nutricionais e na composição mineral em plantas de camucamuzeiro, conduziu-se experimento em casa de vegetação, mediante a técnica do elemento faltante. O delineamento experimental foi o inteiramente casualizado, com quatro repetições e oito tratamentos, sendo completo (N, P, K, Ca, Mg, S e micronutrientes) e omissão individual de N, P, K, Ca, Mg, S e B. Os sintomas visuais de deficiências foram, de modo geral, de fácil caracterização para todos os nutrientes. Com exceção do fósforo, as omissões dos demais nutrientes afetaram a produção de matéria seca, quando comparados ao tratamento completo. Com base nos teores em g kg-1, dos macronutrientes, e em mg kg-1, do micronutriente boro nas folhas, infere-se em uma primeira aproximação dos valores adequados (completo), ou seja: 16,9 a 18,2 de N ; 1,2 a 1,9 de P; 5,2 a 6,0 de K; 9,9 a 11,7 de Ca; 1,4 a 3,6 de Mg; 2,4 a 2,8 de S ; 8,4 a 9,5 de B e do deficiente (omissão) , 6,5 a 7,9 de N ; =0,9 de P; =1,7 de K ; 5,4 a 6,5 de Ca; =0,7 de Mg; 0,7 a 1,2 de S e 1,1 a 1,9 de B

Tratamentos pós-colheita com 1-metilciclopropeno em manga 'Tommy Atkins': efeito de doses e número de aplicações

Foram conduzidos dois experimentos com o objetivo de avaliar a eficiência de tratamentos com 1-MCP sobre a conservação da manga 'Tommy Atkins', reconhecendo doses e número de aplicações. Os frutos foram colhidos no estádio de maturação 2, em pomar comercial localizado em Petrolina, Pernambuco. No primeiro experimento, foram testados: 1. doses (0; 600; 1.200 e 2.400 nL L-1), aplicadas a 25ºC; e 2. tempo de armazenamento (0; 2; 4; 7; 9; 10 e 11 dias), em temperatura ambiente (26,7±2,0ºC). O delineamento experimental foi o inteiramente casualizado, em fatorial 4 x 7 (doses de 1-MCP x tempo de armazenamento), com 4 repetições. No segundo experimento, foram estudados: 1. doses e número de aplicações (controle, uma aplicação de 900 nL L-1, uma aplicação de 1.200 nL L-1 e duas aplicações de 900 nL L-1); e 2. tempo de armazenamento (0; 7; 15; 18; 20; 22; 25 e 26 dias), sendo os frutos mantidos refrigerados (11,0±1,6ºC e 88±7% UR) até o 15º dia e, então, transferidos para temperatura ambiente (26,3±2,1ºC e 44±6% UR). Neste experimento, o delineamento foi inteiramente casualizado, em fatorial 4 x 8 (dose e número de aplicações de 1-MCP x tempo de armazenamento), com 4 repetições. A primeira ou única aplicação ocorreu no dia da colheita e a segunda, no 14º dia. O 1-MCP afetou essencialmente a firmeza da polpa, sendo 1.200 nL L-1 a dose mais eficiente no retardo do amaciamento. A realização de uma aplicação de 1.200 nL L-1 ou duas de 900 nL L-1, sob refrigeração, resultou em efeitos praticamente equivalentes. Porém, as alterações logísticas promovidas por uma única aplicação são menores.

Evaluación de un artículo científico

El proceso de revisión y evaluación de manuscritos, conocido como arbitraje científico o peer review, supone actualmente un paso que se considera necesario para la divulgación de los avances científicos de una determinada disciplina. Cuando una revista recibe un artículo, sus editores recurren a revisores expertos que, después de la lectura detenida del manuscrito, evalúan la idoneidad de su publicación, en este trabajo se describe el proceso editorial de un artículo y se muestran los ítems por los cuales se evalúan los artículos.

Caracterizacion quimica de la vidriera del roseton del Duomo de Siena (Italia, 1288-1289)

Se han caracterizado químicamente una colección de vidrios arquitectónicos coloreados originales del rosetón del Duomo (catedral) de Siena, Italia, realizados bajo diseño del artista Duccio di Buoninsegna en 1288-89. Esta comunicación expone los resultados obtenidos mediante el empleo de microsonda electrónica de Castaing (mayoritarios) y espectrometría de masas con fuente de plasma acoplado inductivamente (ICP-MS, elementos en traza) en vidrios de varios colores (incoloro, verde oscuro, verde oliva, amarillo, violeta, rosa, azul oscuro, azul celeste, rojo plaqué). Se trata de vidrios sódico-cálcicos (valores en peso alrededor del 13-14 % de Na2O, 56-64 % SiO2, 4% MgO, 9-10 % CaO, 2,5-4 K2O) de tradición por tanto mediterránea. Un resultado semejante se encontró precedentemente para vidrios realizados a lo largo del siglo XIV, cuanto menos unos 40 años después (vidriera de la iglesia del Monestir de Pedralbes, Barcelona). En general, el estado de conservación de estos vidrios es bueno, excepto por lo que se refiere a la adherencia de las grisallas al vidrio base.

Dépistage du cancer ovarien dans la population générale [Ovarian cancer screening in the general population].

Although the incidence of ovarian cancer is low, mortality from this cancer is high due to discovery at a late stage in the majority of cases. So it seems worthwhile to detect ovarian cancer at an early stage. The clinical presentation is nonspecific, thus screening tools have been evaluated. The most efficient screening technique includes two steps: evaluation of CA-125 and then sonography in case of abnormal results of CA-125. Two main studies have been performed in large populations. The PLCO-study has led to negative results: no reduction in ovarian cancer mortality in the screening group with an important increase in surgical morbidity. The final results of the UKCTOCS-study will be known in two years. Currently these data can't allow the realization of ovarian cancer screening in the general population, mainly due to their natural history.

Glutamine and antioxidants in the critically ill patient: a post hoc analysis of a large-scale randomized trial.

BACKGROUND: The recent large randomized controlled trial of glutamine and antioxidant supplementation suggested that high-dose glutamine is associated with increased mortality in critically ill patients with multiorgan failure. The objectives of the present analyses were to reevaluate the effect of supplementation after controlling for baseline covariates and to identify potentially important subgroup effects. MATERIALS AND METHODS: This study was a post hoc analysis of a prospective factorial 2 × 2 randomized trial conducted in 40 intensive care units in North America and Europe. In total, 1223 mechanically ventilated adult patients with multiorgan failure were randomized to receive glutamine, antioxidants, both glutamine and antioxidants, or placebo administered separate from artificial nutrition. We compared each of the 3 active treatment arms (glutamine alone, antioxidants alone, and glutamine + antioxidants) with placebo on 28-day mortality. Post hoc, treatment effects were examined within subgroups defined by baseline patient characteristics. Logistic regression was used to estimate treatment effects within subgroups after adjustment for baseline covariates and to identify treatment-by-subgroup interactions (effect modification). RESULTS: The 28-day mortality rates in the placebo, glutamine, antioxidant, and combination arms were 25%, 32%, 29%, and 33%, respectively. After adjusting for prespecified baseline covariates, the adjusted odds ratio of 28-day mortality vs placebo was 1.5 (95% confidence interval, 1.0-2.1, P = .05), 1.2 (0.8-1.8, P = .40), and 1.4 (0.9-2.0, P = .09) for glutamine, antioxidant, and glutamine plus antioxidant arms, respectively. In the post hoc subgroup analysis, both glutamine and antioxidants appeared most harmful in patients with baseline renal dysfunction. No subgroups suggested reduced mortality with supplements. CONCLUSIONS: After adjustment for baseline covariates, early provision of high-dose glutamine administered separately from artificial nutrition was not beneficial and may be associated with increased mortality in critically ill patients with multiorgan failure. For both glutamine and antioxidants, the greatest potential for harm was observed in patients with multiorgan failure that included renal dysfunction upon study enrollment.

Metabolic effects of insulin and IGFs on gilthead sea bream (Sparus aurata) muscle cells.

Primary cultures of gilthead sea bream myocytes were performed in order to examine the relative metabolic function of insulin compared with IGF-I and IGF-II (insulin-like growth factors, IGFs) at different stages in the cell culture. In these cells, the in vitro effects of insulin and IGFs on 2-deoxyglucose (2-DG) and L-alanine uptake were studied in both myocytes (day 4) and small myotubes (day 9). 2-DG uptake in gilthead sea bream muscle cells was increased in the presence of insulin and IGFs in a time dependent manner and along with muscle cell differentiation. On the contrary, L-alanine uptake was also stimulated by insulin and IGFs but showed an inverse pattern, being the uptake higher in small myocytes than in large myotubes. The results of preincubation with inhibitors (PD-98059, wortmannin, and cytochalasin B) on 2-DG uptake indicated that insulin and IGFs stimulate glucose uptake through the same mechanisms, and evidenced that mitogenesis activator protein kinase (MAPK) and PI3K-Akt transduction pathways mediate the metabolic function of these peptides. In the same way, we observed that GLUT4 protein synthesis was stimulated in the presence of insulin and IGFs in gilthead sea bream muscle cells in a different manner at days 4 or 9 of the culture. In summary we describe here, for the first time, the effects of insulin and IGFs on 2-DG and L-alanine uptake in primary culture of gilthead sea bream muscle cells. We show that both MAPK and PI3K-Akt transduction pathways are needed in order to control insulin and IGFs actions in these cells. Moreover, changes in glucose uptake can be explained by the action of the GLUT4 transporter, which is stimulated in the presence of insulin and IGFs throughout the cell culture.

Efeito da concentração de sacarose na germinação in vitro do pólen de cinco acessos de bacurizeiro (Platonia insignis MART.)

O bacurizeiro é uma das espécies fruteiras nativas de maior importância socioeconômica das regiões Norte e Meio-Norte do Brasil. No entanto, ainda há carência de conhecimentos científicos sobre a espécie, especialmente sobre sua biologia floral, mecanismos reprodutivos e viabilidade do pólen. O conhecimento sobre a viabilidade do pólen é fundamental para o melhoramento genético, especialmente quando se tem em mente a realização de hibridizações controladas. O objetivo deste trabalho foi avaliar o efeito da sacarose e de acessos na viabilidade do pólen de bacurizeiro por meio da germinação in vitro. Avaliaram-se cinco acessos (BGB 6, BGB 11, BGB 48, BGB 32 e BGB 16) de bacurizeiro combinados com um tratamento-controle, sem sacarose, e quatro concentrações de sacarose (5; 7,5; 10 e 20%), em delineamento inteiramente ao acaso, em arranjo fatorial 5 x 5, com oito repetições. Na maioria dos acessos, a germinação in vitro do pólen atingiu um máximo (71,1 e 63,0%) entre as concentrações de sacarose de 9,4 e 9,7% e, a partir daí, caiu de forma drástica até atingir um percentual inferior a 10% de germinação na concentração de 20% de sacarose. Foram encontrados bons percentuais de germinação (entre 63,0 e 77,1%) para todos os acessos, com exceção do acesso BGB 16, que não alcançou 50%.

Tumour biology, metastatic sites and taxanes sensitivity as determinants of eribulin mesylate efficacy in breast cancer: results from the ERIBEX retrospective, international, multicenter study.

BACKGROUND: Our retrospective, international study aimed at evaluating the activity and safety of eribulin mesylate (EM) in pretreated metastatic breast cancer (MBC) in a routine clinical setting. METHODS: Patients treated with EM for a locally advanced or MBC between March 2011 and January 2014 were included in the study. Clinical and biological assessment of toxicity was performed at each visit. Tumour response was assessed every 3 cycles of treatment. A database was created to collect clinical, pathological and treatment data. RESULTS: Two hundred and fifty-eight patients were included in the study. Median age was 59 years old. Tumours were Hormone Receptor (HR)-positive (73.3 %) HER2-positive (10.2 %), and triple negative (TN, 22.5 %). 86.4 % of the patients presented with visceral metastases, mainly in the liver (67.4 %). Median previous metastatic chemotherapies number was 4 [1-9]. Previous treatments included anthracyclines and/or taxanes (100 %) and capecitabine (90.7 %). Median number of EM cycles was 5 [1-19]. The relative dose intensity was 0.917. At the time of analysis (median follow-up of 13.9 months), 42.3 % of the patients were still alive. The objective response rate was 25.2 % (95 %CI: 20-31) with a 36.1 % clinical benefit rate (CBR). Median time to progression (TTP) and overall survival were 3.97 (95 %CI: 3.25-4.3) and 11.2 (95 %CI: 9.3-12.1) months, respectively. One- and 2-year survival rates were 45.5 and 8.5 %, respectively. In multivariate analysis, HER2 positivity (HR = 0.29), the presence of lung metastases (HR = 2.49) and primary taxanes resistance (HR = 2.36) were the only three independent CBR predictive factors, while HR positivity (HR = 0.67), the presence of lung metastases (HR = 1.52) and primary taxanes resistance (HR = 1.50) were the only three TTP independent prognostic factors. Treatment was globally well tolerated. Most common grade 3-4 toxicities were neutropenia (20.9 %), peripheral neuropathy (3.9 %), anaemia (1.6 %), liver dysfunction (0.8 %) and thrombocytopenia (0.4 %). Thirteen patients (5 %) developed febrile neutropenia. CONCLUSION: EM is an effective new option in heavily pretreated MBC, with a favourable efficacy/safety ratio in a clinical practice setting. Our results comfort the use of this new molecule and pledge for the evaluation of EM-trastuzumab combination in this setting. Tumour biology, primary taxanes sensitivity and metastatic sites could represent useful predictive and prognostic factors.

CTP in Transient Global Amnesia: A Single-Center Experience of 30 Patients.

BACKGROUND AND PURPOSE: Medial temporal lobe abnormalities on DWI and functional imaging are occasionally observed in patients with transient global amnesia. We used CTP to study these patients during or briefly after resolution of their amnesic syndrome. MATERIALS AND METHODS: From 2002 onward, patients satisfying clinical criteria for transient global amnesia who underwent CTP were included. Patients with additional clinical features suggesting transient ischemic attack or stroke and those with an ischemic lesion on subsequent DWI were excluded. If deemed necessary by the clinician, DWI was performed within 10 days. RESULTS: Thirty patients with transient global amnesia underwent CTP at a median latency of 5.9 hours (interquartile range, 4.3-9.7 hours) after symptom onset. All findings, except for those in 1 patient, were normal, including those in the 14 patients with well-imaged hippocampi. In the patient with abnormal findings, CTP and PWI showed hypoperfusion in both lentiform nuclei extending into the insulae, with normalization on the repeat CTP 6 days later. In 10 patients, DWI was performed at a median latency of 2 days (interquartile range, 0-9 days). Of these, 2 showed punctate hippocampal lesions, often seen in transient global amnesia. In 2 patients excluded because of mildly atypical transient global amnesia and ischemic lesions on subsequent DWI, acute CTP findings were also normal. CONCLUSIONS: Patients with transient global amnesia had normal CTP findings in the acute phase with the exception of 1 patient with transient hypoperfusion in both basal ganglia. If imaging is performed for typical and atypical transient global amnesia, DWI should be the preferred method.

Eligibility to treatment and economic effect of the implementation of the new U.S. or European Society of Hypertension/European Society of Cardiology hypertension guidelines.

OBJECTIVE: To evaluate the population and economic impact of implementing the new Joint National Committee (JNC) or European Society of Hypertension (ESH)/European Society of Cardiology (ESC) hypertension guidelines in the Swiss population. METHODS: Cross-sectional, population-based sample (6708 participants) collected between 2003 and 2006 in the city of Lausanne, Switzerland. Blood pressure categories were defined according to both the JNC (JNC-7 and JNC-8) and the ESH/ESC (2007 and 2013) guidelines. RESULTS: The proportion of participants aged 35-60 years eligible for drug treatment was 25.6% [95% confidence interval (CI) 24.4-26.9%] and 24.8% (95% CI 23.6-26.0%) for the JNC-7 and the JNC-8 guidelines, respectively; for participants aged 60-75 years, the values were 62.3% (95% CI 60.1-64.5%) and 46.8% (95% CI 44.5-49.0%), respectively. Shifting from the JNC-7 to the JNC-8 guidelines would lead to an annual saving of 163.6 million Swiss francs (187.7 million US dollars or 134.5 million European euro). The proportion of participants aged 35-75 years without chronic kidney disease, diabetes mellitus or reported history of cardiovascular disease and eligible for treatment was 30.2% (95% CI 29.0-31.4%) for the ESH/ESC 2007 and 2013 guidelines. For participants with chronic kidney disease, diabetes mellitus or reported history of cardiovascular disease, the values were 73.6% (95% CI 70.8-76.3%) and 55.6% (95% CI 52.5-58.8%), respectively. Shifting from the ESH/ESC 2007 to the ESH/ESC 2013 guidelines would lead to an annual saving of 86.9 million Swiss francs (99.5 million US dollars or 71.4 million European euro). CONCLUSION: In Switzerland, shifting from the JNC-7 to the JNC-8 guidelines or from the ESH/ESC 2007 to the ESH/ESC 2013 guidelines would decrease the prevalence of patients eligible for treatment and increase the percentage of treated patients within blood pressure goals. Both strategies lead to potential savings in antihypertensive drug treatment.