RESUMO
The majority of the world's natural rubber comes from the rubber tree (Hevea brasiliensis). As a key enzyme for synthesizing phenylpropanoid compounds, phenylalanine ammonia-lyase (PAL) has a critical role in plant satisfactory growth and environmental adaptation. To clarify the characteristics of rubber tree PAL family genes, a genome-wide characterization of rubber tree PALs was conducted in this study. Eight PAL genes (HbPAL1-HbPAL8), which spread over chromosomes 3, 7, 8, 10, 12, 13, 14, 16, and 18, were found to be present in the genome of H. brasiliensis. Phylogenetic analysis classified HbPALs into groups I and II, and the group I HbPALs (HbPAL1-HbPAL6) displayed similar conserved motif compositions and gene architectures. Tissue expression patterns of HbPALs quantified by quantitative real-time PCR (qPCR) proved that distinct HbPALs exhibited varying tissue expression patterns. The HbPAL promoters contained a plethora of cis-acting elements that responded to hormones and stress, and the qPCR analysis demonstrated that abiotic stressors like cold, drought, salt, and H2O2-induced oxidative stress, as well as hormones like salicylic acid, abscisic acid, ethylene, and methyl jasmonate, controlled the expression of HbPALs. The majority of HbPALs were also regulated by powdery mildew, anthracnose, and Corynespora leaf fall disease infection. In addition, HbPAL1, HbPAL4, and HbPAL7 were significantly up-regulated in the bark of tapping panel dryness rubber trees relative to that of healthy trees. Our results provide a thorough comprehension of the characteristics of HbPAL genes and set the groundwork for further investigation of the biological functions of HbPALs in rubber trees.
Assuntos
Regulação da Expressão Gênica de Plantas , Hevea , Família Multigênica , Fenilalanina Amônia-Liase , Proteínas de Plantas , Perfilação da Expressão Gênica , Genoma de Planta , Hevea/genética , Hevea/enzimologia , Hevea/metabolismo , Fenilalanina Amônia-Liase/genética , Fenilalanina Amônia-Liase/metabolismo , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas/genética , Estresse Fisiológico/genéticaRESUMO
Glyphosate application, even in low doses, changes the metabolism of crops. This research aimed to evaluate the effects of glyphosate low doses and sowing season on metabolic changes of early-cycle common beans. Two experiments were conducted in the field, one in the winter season and one in the wet season. The experimental design was a randomized complete block design consisting of the application of glyphosate low doses [0.0, 1.8, 7.2, 12.0, 36.0, 54.0, and 108.0 g acid equivalent (a.e.) ha-1] in the phenological stage V4 with four replications. In the winter season, glyphosate and shikimic acid were increased five days after the application of treatments. In contrast, the same compounds increased only at doses of 36 g a.e. ha-1 and above in the wet season. The dose of 7.2 g a.e. ha-1 increased phenylalanine ammonia-lyase and benzoic acid in the winter season. The doses of 54 and 108 g a.e. ha-1 increased benzoic acid, caffeic acid, and salicylic acid. Our study indicated that glyphosate low doses increase the concentration of shikimic, benzoic, salicylic and caffeic acid, PAL and tyrosine. There was no reduction in aromatic amino acids and in secondary compounds from the shikimic acid pathway.
Assuntos
Herbicidas , Phaseolus , Benzoatos , Herbicidas/farmacologia , Phaseolus/metabolismo , Metabolismo Secundário , Ácido Chiquímico/metabolismo , GlifosatoRESUMO
In this study, different plasma-activated liquids were evaluated for their antimicrobial effects against Escherichia coli, as well as for their cytotoxicity on mammalian cells. The PALs were prepared from distilled (DIS), deionized (DI), filtered (FIL), and tap (TAP) water. Additionally, 0.9% NaCl saline solution (SAL) was plasma-activated. These PALs were prepared using 5 L/min air gliding arc plasma jet for up to 60.0 min of exposure. Subsequently, the physicochemical properties, such as, the oxidation-reduction potential (ORP), the pH, the conductivity, and the total dissolved solids (TDS) were characterized by a water multiparameter. The PALs obtained showed a drastic decrease in the pH with increasing plasma exposure time, in contrast, the conductivity and TDS increased. In a general trend, the UV-vis analyses identified a higher production of the following reactive species of nitrogen and oxygen (RONS), HNO2, H2O2, NO3-, and NO2-. Except for the plasma-activated filtered water (PAW-FIL), where there was a change in the position of NO2- and NO3- at some pHs, The higher production of HNO2 and H2O2-reactive species was observed at a low pH. Finally, the standardized suspensions of Escherichia coli were exposed to PAL for up to 60.0 min. The plasma-activated deionized water (PAW-DI pH 2.5), plasma-activated distilled water (PAW-DIS pH 2.5 and 3), and plasma-activated tap water (PAW-TAP 3.5) showed the best antimicrobial effects at exposure times of 3.0, 10.0, and 30.0 min, respectively. The MTT analysis demonstrated low toxicity of all of the PAL samples. Our results indicate that the plasma activation of different liquids using the gliding arc system can generate specific physicochemical conditions that produce excellent antibacterial effects for E. coli with a safe application, thus bringing future contributions to creating new antimicrobial protocols.
Assuntos
Anti-Infecciosos , Gases em Plasma , Animais , Antibacterianos/farmacologia , Escherichia coli , Peróxido de Hidrogênio/química , Mamíferos , Dióxido de Nitrogênio , Gases em Plasma/farmacologia , Gases em Plasma/química , Água/químicaRESUMO
Broccoli contains a high content of nutraceutical compounds, such as glucosinolates and flavonoids. In this work, the effect of different treatments that modulate postharvest senescence of broccoli was evaluated and flavonoid metabolism during postharvest storage was analyzed at 20°C. A decrease in hue angle (HUE°) and chlorophylls and an increase in flavonoid content were detected during senescence. It observed that most of the treatments that delayed senescence also decreased flavonoid content, except visible light and UV-C treatments. In all cases, a direct correlation between those treatments that increased flavonoid biosynthesis and BoPAL gene expression was detected. This response was not detected in the expression of the other two flavonoid synthesis relevant genes BoCHS and BoCHI, suggesting that BoPAL has a greater influence on the regulation of the via, during broccoli senescence. PRACTICAL APPLICATIONS: Broccoli is a vegetable with valuable nutritional properties. Because it is in full development at the time of harvest, it has a short shelf life. In this work, it is showed that visible light and UV-C treatments not only delayed the senescence of broccoli, but also increased flavonoid content. Our results suggest that the most important enzyme in the phenylpropanoid biosynthesis pathway during broccoli postharvest is phenylalanine ammonia lyase, and that this may be a key point in regulating the biosynthesis of these nutritionally valuable compounds.
Assuntos
Brassica , Aciltransferases , Brassica/genética , Brassica/metabolismo , Flavonoides/metabolismo , Liases Intramoleculares , Fenilalanina Amônia-Liase/genética , Fenilalanina Amônia-Liase/metabolismoRESUMO
The production of 3-indoleacetic acid (IAA) by plant growth-promoting bacteria (PGPR) stimulates root development and plant growth. In addition, morphological changes such as an increased root ramification and root hair production improves nutrient absorption and biomass accumulation. The objective of this work was to evaluate the effect of IAA-producing strains on rice in an advanced stage of its vegetative cycle. Rice was inoculated with Gluconacetobacter diazotrophicus PAL 5 and its lao- mutant, deficient in auxin production, Azospirillum baldaniorum Sp 245, and Escherichia coli DH10b. Both the mutant and wild-type G. diazotrophicus stimulated root elongation, area, volume, and diameter. However, the lao- mutant strain was the only one capable of increasing the number of roots. In turn, inoculation with A. baldaniorum had no significant effect on plant development. The inoculation with E. coli led to changes in root volume, area, and diameter, and a response that may be related to the stress caused by its presence. We conclude that the inoculation with G. diazotrophicus stimulates the root system's growth independently of their IAA production ability, suggesting that a metabolite other than IAA is responsible for this effect at advanced stages of the rice's vegetative cycle.
Assuntos
Oryza , Escherichia coli/genética , Escherichia coli/metabolismo , Ácidos Indolacéticos/metabolismo , Oryza/microbiologia , Raízes de Plantas/microbiologiaRESUMO
Raspberries are important sources of bioactive compounds, whose synthesis is influenced by the fertilization system and the maturity stage. This study evaluated the effect of organic and conventional fertilization systems on raspberries at three maturity stages, pink, ripe, and overripe. Physicochemical characteristics, bioactive compounds (phenolic profile, vitamin C), antioxidant capacity (DPPH, FRAP, TEAC, and ORAC), phenolic-associated enzyme, phenylalanine ammonia lyase (PAL), and antioxidant enzymes (SOD, CAT, GPx, and APX) were evaluated. The physicochemical determination of the fruit did not reveal differences between fertilization systems. Regarding bioactive compounds, higher content of anthocyanins was found in organic raspberries at all maturity stages. Organic fertilization increased the content of ellagic acid and gallic acid at all stages of maturity. Higher content of caffeic, hydroxybenzoic, protocatechuic acid, and vitamin C was observed in organic raspberry at the overripe stage. Raspberries grown with organic fertilization exhibited higher values of antioxidant capacity by the DPPH, FRAP, and TEAC methods at all maturity stages. Raspberries under organic fertilization showed significantly greater activity of CAT, SOD, APX, GPX, and PAL. The present study suggests that organic fertilization induces oxidative stress causing an increase in antioxidant defense mechanisms, enhancing bioactive compound production, and improving antioxidant capacity in raspberries.
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This study evaluated the chemical compositions of the leaves and fruits of eight black pepper cultivars cultivated in Pará State (Amazon, Brazil). Hydrodistillation and gas chromatography-mass spectrometry were employed to extract and analyze the volatile compounds, respectively. Sesquiterpene hydrocarbons were predominant (58.5-90.9%) in the cultivars "Cingapura", "Equador", "Guajarina", "Iaçará", and "Kottanadan", and "Bragantina", "Clonada", and "Uthirankota" displayed oxygenated sesquiterpenoids (50.6-75.0%). The multivariate statistical analysis applied using volatile composition grouped the samples into four groups: γ-Elemene, curzerene, and δ-elemene ("Equador"/"Guajarina", I); δ-elemene ("Iaçará"/"Kottanadan"/"Cingapura", II); elemol ("Clonada"/"Uthirankota", III) and α-muurolol, bicyclogermacrene, and cubebol ("Bragantina", IV). The major compounds in all fruit samples were monoterpene hydrocarbons such as α-pinene, ß-pinene, and limonene. Among the cultivar leaves, phenolics content (44.75-140.53 mg GAE·g-1 FW), the enzymatic activity of phenylalanine-ammonia lyase (20.19-57.22 µU·mL-1), and carotenoids (0.21-2.31 µg·mL-1) displayed significant variations. Due to black pepper's susceptibility to Fusarium infection, a molecular docking analysis was carried out on Fusarium protein targets using each cultivar's volatile components. F. oxysporum endoglucanase was identified as the preferential protein target of the compounds. These results can be used to identify chemical markers related to the susceptibility degree of black pepper cultivars to plant diseases prevalent in Pará State.
Assuntos
Piper nigrum/metabolismo , Sesquiterpenos/análise , Sesquiterpenos/metabolismo , Brasil , Frutas/química , Frutas/genética , Cromatografia Gasosa-Espectrometria de Massas/métodos , Metaboloma , Simulação de Acoplamento Molecular , Monoterpenos/análise , Monoterpenos/metabolismo , Óleos Voláteis/química , Piper nigrum/genética , Folhas de Planta/genética , Óleos de Plantas/química , Sesquiterpenos/químicaRESUMO
BACKGROUND: Accumulation of phenylalanine (Phe) due to deficiency in the enzyme phenylalanine hydroxylase (PAH), responsible for the conversion of Phe into tyrosine leads to Phenylketonuria (PKU), a rare autosomal recessive inborn error of metabolism with a mean prevalence of approximately 1:10,000 to 1:15,000 newborns. Physical, neurocognitive and psychiatric symptoms include neurodevelopmental disorder as intellectual disability and autism spectrum disorder. The most common treatments such as low-Phe diet and supplements may decrease blood Phe concentrations, but neuropsychological, behavioral and social issues still occur in some patients. This study aimed to better understand (i) the Brazilian population's knowledge about newborn screening (NBS), the main diagnostic method for PKU, as well as (ii) the impacts of phenylketonuria in the daily lives of patients and parents. METHODS: Two surveys in Real World Data format gathering of Brazilian residents by online questionnaires with (i) 1000 parents of children up to 5 years old between March and April 2019; (ii) 228 PKU patients and caregivers in March 2019. The survey was conducted in partnership with Abril Publisher and two Brazilian patient associations: Metabolic Mothers and SAFE Brasil, for families with rare diseases and PKU patients, respectively. RESULTS: The first questionnaire shows that 93% of parents recognize the importance of NBS and 92% report that their children have undergone the test. Still, two out of ten participants did not know what the exam is or what it is for. From the second questionnaire nine out of ten patients had their PKU diagnosis by NBS. Although strict dietary controls for PKU were claimed by 44% of respondents from second questionnaire, 55% assume not following all nutritionist recommendations and 52% did not maintain routinely Phe control levels. In addition, 53% said they had high spending on medical appointments, therapies and purchase of special foods. CONCLUSIONS: Despite the lack of understanding, the awareness of NBS importance is present in the studied population. The early diagnosis of most PKU patients in the study corroborates with neonatal screening central role of PKU early detection. The difficulty in adhering to dietary adjustments and the possibility that current and new therapeutic strategies other than diet could be determinant to achieve the recommended Phe levels.
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BACKGROUND: The assessment of patients' quality of life (QOL) is essential when evaluating the outcome of palliative care; however, no instruments have been validated for measuring symptoms and QOL in patients receiving palliative care in Chile. We aimed to investigate the content validity of the EORTC Quality of Life Questionnaire Core 15 Palliative Care (QLQ-C15-PAL), replicating the methods used previously to shorten the EORTC Quality of Life Questionnaire Core 30 (QLQ-C30) for use among patients in palliative care. METHODS: This cross-sectional study was conducted from October to November 2017 in four palliative care services. Patients with advanced cancer and health care professionals (HCPs) were invited to individual interviews to determine the relevance, appropriateness and relative importance of the 30 items of the QLQ-C30 for evaluating the outcome of palliative care, and whether relevant additional issues should be included. RESULTS: A total of 48 patients and 35 HCPs participated. The most important dimensions selected were pain, physical functioning, sleeping difficulties, emotional functioning, nausea/vomiting, fatigue, social functioning, lack of appetite, role functioning and constipation. Qualitative data identified important additional issues not covered by the questionnaire such as satisfaction with care, emotions and psychological support, as well as linguistic issues in the dyspnea and constipation items. CONCLUSIONS: The EORTC QLQ-C15-PAL showed good content validity in the assessment of symptoms and QOL of advanced cancer patients; therefore, we recommend the use of this questionnaire in palliative care in Chile. Dyspnea and constipation items were revised by the EORTC group. More research is needed to add a social dimension for a comprehensive assessment of patients' QOL.
Assuntos
Neoplasias/complicações , Psicometria/normas , Qualidade de Vida/psicologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Chile , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/psicologia , Cuidados Paliativos , Psicometria/instrumentação , Psicometria/métodos , Inquéritos e QuestionáriosRESUMO
Introduction: Due to inherent limitations of chlorhexidine, search for an effective and potentially safe anti-plaque agent has led to emergence of alternative products. Aim: To evaluate and compare the effects of Oral Pal Plus mouth rinse and chlorhexidine on dental plaque and gingivitis. Materials and Methods: The present study was randomized parallel group controlled trial. A group of 90 healthy subjects in the age group of 18-21 yrs received complete supragingival scaling at baseline and study variables viz, Plaque index and Gingival index were recorded. Subjects were then randomly divided into three groups (30 in each group) and were randomly intervened with three different mouthwashes. ie, Chlorhexidine, Oral Pal Plus and normal saline. Variables were again recorded on the 7th day 14th day after use of mouthwashes and data obtained was subjected to statistical analysis. Results: There was statistically significant reduction in plaque and gingival scores from baseline to 14 days in both the groups A & B. Conclusion: Chlorhexidine (0.2%) and Oral Pal Plus mouthwash showed significantly reduction in plaque scores and gingival scores whereas no improvement was seen in Group C using normal saline over 14 days. (AU)
Assuntos
Clorexidina , Ensaio Clínico Controlado Aleatório , Placa Dentária , GengiviteRESUMO
The present study evaluated the effects of ultraviolet B (UVB) radiation and wounding stress, applied alone or combined, on the biosynthesis of phenolic compounds and ascorbic acid in the peel and pulp of red prickly pear (Opuntia ficus-indica cv. Rojo Vigor). Whole and wounded-fruit samples were treated with UVB radiation (6.4 W·m-2) for 0 and 15 min, and stored for 24 h at 16 °C. Phytochemical analyses were performed separately in the peel and pulp. The highest phenolic accumulation occurred after storage of the whole tissue treated with UVB, where the main phenolic compounds accumulated in the peel and pulp were quercetin, sinapic acid, kaempferol, rosmarinic acid, and sinapoyl malate, showing increases of 709.8%, 570.2%, 442.8%, 439.9%, and 186.2%, respectively, as compared with the control before storage. Phenylalanine ammonia-lyase (PAL) activity was increased after storage of the whole and wounded tissue treated with UVB light, and this increase in PAL activity was associated to phenolic accumulation. On the other hand, l-galactono-γ-lactone dehydrogenase (GalLDH) activity and ascorbic acid biosynthesis was enhanced due to UVB radiation, and the effect was increased when UVB was applied in the wounded tissue showing 125.1% and 94.1% higher vitamin C content after storage when compared with the control. Respiration rate was increased due to wounding stress, whereas ethylene production was increased by wounding and UVB radiation in prickly pears. Results allowed the generation of a physiological model explaining the UVB and wound-induced accumulation of phenolic compounds and ascorbic acid in prickly pears, where wounding facilitates UVB to access the underlying tissue and enhances an apparent synergistic response.
Assuntos
Modelos Biológicos , Opuntia/metabolismo , Fenóis/metabolismo , Proteínas de Plantas/metabolismo , Raios UltravioletaRESUMO
Studies aiming at heterologous expression of highly hydrophobic proteins, such as outer membrane proteins in general and peptidoglycan-associated lipoprotein (PAL) in particular, are not trivial due to difficulties in obtaining recombinant protein in a soluble state, which is desired because it allows purification by traditional chromatographic methods. PAL is associated with the integrity of the cellular envelope in Gram-negative bacteria and interacts strongly with the peptidoglycan layer. However, it is incorporated into inclusion bodies in studies focusing on its heterologous production. This protocol describes an efficient protein refolding method to solubilize and purify a recombinant PAL. Initially, recombinant PAL-enriched inclusion bodies obtained after the induction of PAL expression in Escherichia coli are treated with 8 M urea and then undergo buffer exchange via dialysis. Afterward, the soluble, recombinant PAL is purified using standard chromatographic methods. © 2018 by John Wiley & Sons, Inc.
Assuntos
Proteínas de Bactérias , Escherichia coli , Expressão Gênica , Lipoproteínas , Dobramento de Proteína , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Escherichia coli/química , Escherichia coli/genética , Escherichia coli/metabolismo , Lipoproteínas/biossíntese , Lipoproteínas/química , Lipoproteínas/genética , Lipoproteínas/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
We investigated the effects of high-Ca fat-free milk phase (MD) (prescription of approximately 1500 mg of Ca/d) v. low-Ca phase (CD) (prescription of approximately 800 mg of Ca/d) in an energy-restricted diet on the metabolic syndrome (MetS) and cardiometabolic measures in individuals with type 2 diabetes mellitus (T2DM) and low habitual Ca consumption (<600 mg/d). In this randomised cross-over design, fourteen adults with T2DM (49·5 (sd 8·6) years, BMI 29·4 (sd 4·5) kg/m2) consumed either MD or CD for 12 weeks, with a washout of 18 weeks between phases. A breakfast shake containing 700 mg (MD) or 6·4 mg (CD) of Ca was consumed in the laboratory. In addition, energy-restricted diets were prescribed (800 mg of dietary Ca/d). Waist circumference (WC), fasting glucose, fasting TAG, systolic (SBP) and diastolic blood pressure (DBP), fasting total cholesterol, fasting LDL-cholesterol, fasting HDL-cholesterol, HDL:LDL ratio, HDL:TAG ratio and lipid accumulation product (LAP) index were assessed at baseline and after each phase. Ca consumption during the study was equivalent to 1200 mg/d during MD and 525 mg/d during CD. There was a greater reduction in WC, SBP, DBP and LAP index after MD compared with CD. HDL:LDL ratio increased and total cholesterol, LDL-cholesterol, SBP, DBP and LAP index decreased only in MD. The consumption of approximately 1200 mg of Ca/d (700 mg from fat-free milk+500mg from other dietary sources) associated with an energy-restricted diet decreased some of the MetS components and cardiometabolic measures in adults with T2DM.
Assuntos
Cálcio da Dieta/administração & dosagem , Restrição Calórica , Diabetes Mellitus Tipo 2/tratamento farmacológico , Dieta , Comportamento Alimentar , Síndrome Metabólica/tratamento farmacológico , Leite/química , Adulto , Animais , Pressão Sanguínea/efeitos dos fármacos , Cálcio da Dieta/farmacologia , Cálcio da Dieta/uso terapêutico , Colesterol/sangue , Estudos Cross-Over , Diabetes Mellitus Tipo 2/metabolismo , Gorduras na Dieta/administração & dosagem , Feminino , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Circunferência da Cintura/efeitos dos fármacosRESUMO
The Pisum arvense lectin (PAL), a legume protein belonging to the Vicieae tribe, is capable of specific recognition of mannose, glucose and its derivatives without altering its structure. In this work, the three-dimensional structure of PAL was determined by X-ray crystallography and studied in detail by a combination of molecular docking and molecular dynamics (MD). Crystals belonging to monoclinic space group P21 were grown by the vapor diffusion method at 293 K. The structure was solved at 2.16 Å and was similar to that of other Vicieae lectins. The structure presented Rfactor and Rfree of 17.04% and 22.08%, respectively, with all acceptable geometric parameters. Molecular docking was performed to analyze interactions of the lectin with monosaccharides, disaccharides and high-mannose N-glycans. PAL demonstrated different affinities on carbohydrates, depending on bond orientation and glycosidic linkage present in ligands. Furthermore, the lectin interacted with representative N-glycans in a manner consistent with the biological effects described for Vicieae lectins. Carbohydrate-recognition domain (CRD) in-depth analysis was performed by MD, describing the behavior of CRD residues in complex with ligand, stability, flexibility of the protein over time, CRD volume and topology. This is a first report of its kind for a lectin of the Vicieae tribe.
Assuntos
Fabaceae/química , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Lectinas de Plantas/química , Polissacarídeos/química , Cristalografia por Raios XRESUMO
Abstract The phenylalanine (PHE)-restricted diet has improved in quality and diversity over time and has proven to be effective in all patients. Nevertheless, this treatment imposes a heavy social and economic burden to patient and family and impacts quality of life. Sustained adherence to PHE restriction is difficult to maintain. Moreover, even patients with phenylketonuria (PKU) with normal intelligence quotient (IQ) have lower IQ than matched individuals without PKU and can have deficits in multiple other aspects of neuropsychological function, including cognitive and executive function, working memory. They can also have behavior problems, depression, and low self-esteem. In recent years, alternative treatments for PKU have been developed and their use has been indicated for some patients who are candidates for options besides traditional treatment. Sapropterindihydrochloride, large neutral amino acids, and glycomacropeptide are alternative treatment options in use for selected patients. The aim of this article is to review the current knowledge of these new approaches to PKU treatment.
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El Programa de Acreditación de Laboratorios (PAL) se estableció en 1994 con el objeto de mejorar las prestaciones bioquímicas de los Laboratorios Clínicos de la provincia de Buenos Aires. La estructura organizativa es conforme a normas internacionales y los requisitos exigidos para los laboratorios fueron diseñados en un esquema que tuvo en cuenta los avances en los procesos de gestión de los laboratorios y las realidades regionales del país. Con dicha estrategia el programa extendió sus actividades a lo largo y ancho del país. Las auditorías en terreno son desarrolladas por pares evaluadores que cumplen con estrictas normas éticas. Al presente cuenta con más de 1000 laboratorios acreditados ubicados en 18 de las 24 provincias argentinas y su proyección a futuro apunta a hacerse extensivo a la totalidad del territorio nacional.
The laboratories accreditation program (PAL for its name in Spanish) was established in 1994 in order to improve the services of biochemical laboratories of the Province of Buenos Aires. The organisational structure was based on international standards, and the requirements for the laboratories were designed in a scheme that took into account the advances in the management process of the laboratories and the regional realities in Argentina. With this strategy, the Program spread its activities throughout the country. All the audits are conducted in the site, by peer reviewers that follow strict ethical standards. At the present, PAL has more than 1,000 accredited laboratories located in 18 of the 24 Argentine provinces, and its future projection aims at involving the entire national territory.
O Programa de Credenciamento de Laboratórios (PAL, por suas siglas em espanhol) foi criado em 1994 a fim de melhorar os benefícios bioquímicos dos Laboratórios Clínicos da Província de Buenos Aires. A estrutura organizacional está em conformidade com as normas internacionais, e os requisitos exigidos para os laboratórios foram projetados em um esquema que levou em consideração os avanços nos processos de gestão dos laboratórios e as realidades regionais do país. Com esta estratégia, o programa estendeu suas atividades em todo o país. Auditorias no local são realizadas por avaliadores que cumpre os rigorosos padrões éticos. Atualmente, tem mais de 1.000 laboratórios credenciados localizados em 18 das 24 províncias argentinas e sua projeção futura é encaminhada a se estender por todo o território nacional.
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Serviços de Laboratório Clínico/organização & administração , Laboratórios/estatística & dados numéricos , Argentina , Laboratórios/tendências , Gestão da Qualidade TotalRESUMO
Benzyl alcohol and other benzenoid-derived metabolites of particular importance in plants confer floral and fruity flavors to wines. Among the volatile aroma components in Vitis vinifera grape varieties, benzyl alcohol is present in its free and glycosylated forms. These compounds are considered to originate from grapes only and not from fermentative processes. We have found increased levels of benzyl alcohol in red Tannat wine compared to that in grape juice, suggesting de novo formation of this metabolite during vinification. In this work, we show that benzyl alcohol, benzaldehyde, p-hydroxybenzaldehyde, and p-hydroxybenzyl alcohol are synthesized de novo in the absence of grape-derived precursors by Hanseniaspora vineae. Levels of benzyl alcohol produced by 11 different H. vineae strains were 20-200 times higher than those measured in fermentations with Saccharomyces cerevisiae strains. These results show that H. vineae contributes to flavor diversity by increasing grape variety aroma concentration in a chemically defined medium. Feeding experiments with phenylalanine, tryptophan, tyrosine, p-aminobenzoic acid, and ammonium in an artificial medium were tested to evaluate the effect of these compounds either as precursors or as potential pathway regulators for the formation of benzenoid-derived aromas. Genomic analysis shows that the phenylalanine ammonia-lyase (PAL) and tyrosine ammonia lyase (TAL) pathways, used by plants to generate benzyl alcohols from aromatic amino acids, are absent in the H. vineae genome. Consequently, alternative pathways derived from chorismate with mandelate as an intermediate are discussed.
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Benzaldeídos/metabolismo , Álcoois Benzílicos/metabolismo , Aromatizantes/metabolismo , Hanseniaspora/metabolismo , Vitis/microbiologia , Vinho/análise , Benzaldeídos/análise , Álcoois Benzílicos/análise , Fermentação , Aromatizantes/análise , Hanseniaspora/genética , Vitis/metabolismoRESUMO
The intriguing roles of the bacterial Tol-Pal trans-envelope protein complex range from maintenance of cell envelope integrity to potential participation in the process of cell division. In this study, we report the characterization of the XfTolB and XfPal proteins of the Tol-Pal complex of Xylella fastidiosa. X. fastidiosa is a major plant pathogen that forms biofilms inside xylem vessels, triggering the development of diseases in important cultivable plants around the word. Based on functional complementation experiments in Escherichia coli tolB and pal mutant strains, we confirmed the role of xftolB and xfpal in outer membrane integrity. In addition, we observed a dynamic and coordinated protein expression profile during the X. fastidiosa biofilm development process. Using small-angle X-ray scattering (SAXS), the low-resolution structure of the isolated XfTolB-XfPal complex in solution was solved for the first time. Finally, the localization of the XfTolB and XfPal polar ends was visualized via immunofluorescence labeling in vivo during bacterial cell growth. Our results highlight the major role of the components of the cell envelope, particularly the TolB-Pal complex, during the different phases of bacterial biofilm development.
Assuntos
Proteínas da Membrana Bacteriana Externa/química , Biofilmes/crescimento & desenvolvimento , Proteínas de Escherichia coli/química , Regulação Bacteriana da Expressão Gênica , Lipoproteínas/química , Peptidoglicano/química , Proteínas Periplásmicas/química , Xylella/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/metabolismo , Membrana Celular/genética , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Parede Celular/genética , Parede Celular/metabolismo , Parede Celular/ultraestrutura , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Teste de Complementação Genética , Lipoproteínas/genética , Lipoproteínas/metabolismo , Modelos Moleculares , Peptidoglicano/genética , Peptidoglicano/metabolismo , Proteínas Periplásmicas/genética , Proteínas Periplásmicas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Espalhamento a Baixo Ângulo , Homologia de Sequência de Aminoácidos , Difração de Raios X , Xylella/metabolismo , Xylella/ultraestruturaRESUMO
Leaf rust, caused by the foliar pathogen Puccinia triticina is a major disease of wheat in the southern region of Brazil and invariably impacts on production, being responsible for high yield losses. The Brazilian wheat cultivar Toropi has proven, durable adult plant resistance (APR) to leaf rust, which uniquely shows a pre-haustorial resistance phenotype. In this study we aimed to understand the interaction between P. triticina and the pre-haustorial APR in Toropi by quantitatively evaluating the temporal transcription profiles of selected genes known to be related to infection and defense in wheat. The expression profiles of 15 selected genes varied over time, grouping into six expression profile groups. The expression profiles indicated the induction of classical defence pathways in response to pathogen development, but also the potential modification of Toropi's cellular status for the benefit of the pathogen. Classical defence genes, including peroxidases, ß-1,3-glucanases and an endochitinase were expressed both early (pre-haustorial) and late (post-haustorial) over the 72 h infection time course, while induction of transcription of other infection-related genes with a potential role in defence, although variable was maintained through-out. These genes directly or indirectly had a role in plant lignification, oxidative stress, the regulation of energy supply, water and lipid transport, and cell cycle regulation. The early induction of transcription of defence-related genes supports the pre-haustorial resistance phenotype in Toropi, providing a valuable source of genes controlling leaf rust resistance for wheat breeding.
RESUMO
In this work, 25,806 potentially amplifiable microsatellite loci (PAL) were identified in pejerrey, (Odontesthes humensis), with 21% of dinucleotide, 22% trinucleotide, 37% tetranucleotide, 13% pentanucleotide and 7% hexanucleotide. Of the total loci, 167 were classified as "Best PAL", more likely to be variables in populations. The results show that with a small coverage of the genome it was possible to identify a large number of microsatellite loci...