INTRODUÇÃO: Em pacientes com doença pulmonar obstrutiva crônica a reabilitação pulmonar tem demonstrado ser efetiva sobre diversos aspectos, incluindo a melhora física e emocional nas atividades diárias com conseqüente melhora da qualidade de vida e diminuição da dispnéia. Embora os resultados benéficos da atividade física estejam bem documentados, não se sabe, ao certo, os efeitos do exercício físico sobre a lesão pulmonar em pacientes enfisematosos. OBJETIVO: Verificar o efeito pulmonar de dois protocolos de atividade física (intensidade leve-moderada e alta), em ratos Wistar com enfisema pulmonar induzido pela administração intratraqueal de papaína. MÉTODOS: Os animais foram divididos em 5 grupos. Destes, 3 grupos receberam administração intratraqueal de solução de papaína, sendo que um deles permaneceu sedentário (PS) e os outros dois realizaram atividade física intensa (PHE) ou moderada (PME) por 10 semanas após a instalação do enfisema (40 dias após a instilação). O mesmo aconteceu com o grupo que recebeu solução salina (NaCl 0,9%), sendo que um deles permaneceu sedentário (SS) e o outro realizou atividade física intensa (SHE). Foram realizadas medidas de função pulmonar, medidas morfométricas...

O carrapato Rhipicephalus (Boophilus) microplus é o principal ectoparasita bovino e causa importantes perdas econômicas nas criações de bovino. O controle imunológico é estudado como um método alternativo para seu controle, no entanto, uma vacina eficaz ainda necessita ser desenvolvida. A proteína BYC (Boophilus Yolk Pro-Cathepsin) é uma aspártico proteinase presente no ovo do carrapato e envolvida na embriogênese, já tendo sido testada como imunógeno vacinal. O propósito deste estudo foi avaliar se a inoculação de plasmídeos para expressão em células eucarióticas (BYCr-PC e BYCr-PME) contendo a região codificante para a proteína BYC poderiam gerar uma resposta imune específica. A região codificante da proteína BYC foi amplificada por PCR e clonada em dois vetores de expressão eucariotos (pcDNA3 e pME18Neo). Os clones, BYCr-PC e BYCr-PME foram utilizados para testes de inoculação de camundongos BALB/C por via intramuscular. Os camundongos receberam duas inoculações de 100 μg das construções (BYCr-PC ou BYCr-PME) e os controles negativos receberam somente PBS, pcDNA3 ou pME18Neo. A produção de anticorpos, após a inoculação, foi avaliada por Western Blotting e ELISA, sendo detectados anticorpos contra a proteína BYC nos camundongos inoculados com a construção BYCr-PC. A imunolocalização da proteína BYC nas amostras de músculo no local da inoculação foram realizadas com o monoclonal BrBm5 (anti-BYC). Estes resultados mostraram que a inoculação com o plasmídeo BYCr-PC induz a produção de anticorpos específicos e possibilita testar o uso de uma vacina de DNA como um método alternativo para o controle de carrapatos.; The Rhipicephalus (Boophilus) microplus tick is the major bovine ectoparasite and causes important economical losses on cattle breeding. The immunologic control has been studied as an alternative method for the tick control. However...

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); The guava pectin methylesterase (PME) specific activity and vitamin C were assayed in samples from different phases of guava fruit development. The PME enzyme from guava was extracted with borate-acetate buffer, 50 mol/l, pH 8.0, in the presence of NaCl 0.3 mol/l. The results showed PME optimum activity at pH 9 and 95C, and it is a thermostable enzyme. Guava PME retained 96.8% of activity after 300 min in 90C. Electrophoresis showed that guava PME contained two isoforms, one with 57 kDa molecular mass. The analyses of the different phases of guava maturation showed that ascorbic acid decreases during the maturation process, but PME activity increases with maturation.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); In this work we studied the contents of pectin and protein, pectinmethylesterase (PME) activity, and PME stability in various stages of industrial processing, due to the implication of these values on the quality of the final product. The results of the PME stability at different values of pH showed residual PME activity at alkaline pH (7.0, 8.0, 8.5 and 9.5) and high stability at pH 4.0. These results show that pH treatment is not an efficient method to inactivate the PME enzyme. The presence of residual PME activity in all steps of industrial processing was also verified, showing that PME can change the quality of the pulp during storage.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Processo FAPESP: 11/11574-2; Processo FAPESP: 11/04029-8; Processo FAPESP: 10/168267; Processo FAPESP: 13/00424-5; Pós-graduação em Alimentos e Nutrição - FCFAR; O suco de laranja é o suco de fruta mais popular e mais consumido em todo mundo. O Brasil é o maior produtor e exportador mundial de suco de laranja. O consumo de suco de laranja é estimado em mais de 9 bilhões de litros por ano, representando 47% do consumo mundial de suco. A fim de aumentar a vida de prateleira e a estabilidade do suco de laranja, a tecnologia de alta pressão hidrostática (APH) pode ser utilizada como um método alternativo à pasteurização. Este trabalho teve como objetivo otimizar as condições de processamento do suco de laranja da variedade Pêra Rio por APH, pressão (100- 600 MPa), temperatura (30-60 °C) e tempo (30-360 s), avaliar o efeito das condições do processamento de APH no ácido ascórbico e na atividade antioxidante do suco de laranja e avaliar a qualidade do suco de laranja processado a 520 MPa, 60 °C por 360 s, comparativamente ao suco pasteurizado e não processado. A metodologia de superfície de resposta foi empregada para estimar a redução da atividade da enzima pectinametilesterase (PME)...

O objetivo deste estudo foi avaliar, histometricamente, a utilização das proteínas derivadas da matriz do esmalte (PME), associadas ou não à técnica de regeneração óssea guiada (ROG), no tratamento de deiscências ósseas peri-implantares. Foram utilizados seis cães, os quais tiveram os pré molares (p2, p3, p4) e molar (m1) mandibulares extraídos bilateralmente. Após três meses da extração, foram criadas quatro deiscências ósseas vestibulares bilaterais e instalados quatro implantes. As deiscências foram designadas aleatoriamente aos seguintes grupos: PME, ROG, associação PME + ROG e grupo controle. Após dois meses, nos mesmos animais, mais quatro deiscências foram criadas e mais quatro implantes instalados, sendo tratadas da mesma maneira. Três meses depois da primeira intervenção, os animais foram sacrificados e o material processado. As porcentagens médias obtidas para o parâmetro contato ossolimplante, nos grupos PME, ROG, associação PME + ROG e grupo controle, no período de um mês de pós-operatório foram 15,03:t 23,76; 12,87 :t 9,35; 8,52 :t 6,89; 3,88 :t 5,70, respectivamente (p>0,05). No período de três meses: 18,07:t 13,52; 24,42:t 19,88; 36,11 :t 17,53; 16,96:t 17,77, respectivamente (p>0...

Process stability studies towards temperature and/or pressure on pepper pectin methylesterase (PME) were carried out in different systems (purified form, crude extract, pepper pieces and puree) at pH 5.6. Within the temperature range studied (22–80 °C, 5 min), pepper PME in pure form and crude extract was gradually inactivated showing a biphasic inactivation behaviour, indicating the presence of isoenzymes of different thermostability. Pepper samples heated for 15 min showed a maximum of residual PME activity around 55 °C. Isothermal inactivation of pepper PME in purified form and crude extract at pH 5.6 could be described by a biphasic inactivation model for the temperature range studied (62–76 °C). A stable behaviour towards high-pressure/temperature treatments (400–800 MPa/25–60 °C) was observed for crude extract and purified pepper PME. PME in pepper puree samples revealed to be very pressure stable. Mild temperatures combined with pressure treatments seem to increase the extractability from PME in pepper tissue, probably due to the effect on the cell structure.

Os frutos são alimentos fundamentais na alimentação humana e são largamente consumidos em todo o mundo. Parte do consumo é feito na forma de frutos processados aplicados nos mais diversos produtos alimentares como iogurte, gelados, pastelaria, compotas, sumos etc. O processamento dos frutos provoca uma alteração das propriedades organoléticas especialmente na textura, devido a alterações na parede celular e principalmente na lamela média. As homogalacturonanas, um dos polissacarídeos pécticos presentes na lamela média, quando desesterificadas tem capacidade de formar ligações com catiões de Ca2+ formando assim um gel. Essa desesterificação pode ocorrer através da ação da pectina metilesterase (PME). A atividade da PME em conjunto com a adição de iões de cálcio pode trazer aumento da firmeza aos frutos processados. Neste trabalho foi testado o efeito provocado por uma PME em pedaços de morango. O trabalho focou-se na influência da temperatura de enzimagem ao longo do tempo em termos de firmeza dos pedaços. Foram utilizadas 3 temperaturas de incubação da enzima: 4ºC; 25ºC e 40ºC. Os dados obtidos foram adaptados ao modelo matemático de one-phase association que relaciona a firmeza dos pedaços em função do tempo através de uma constante de velocidade de enzimagem...

Mestrado em Ciências Empresariais; Actualmente, percebe-se que os clientes não procuram apenas um produto, pretendem um atendimento personalizado de acordo com as suas necessidades. O Customer Relationship Management (CRM) surge como resposta às organizações que decidem adoptar estratégias de marketing de relacionamento. Os elevados investimentos na compra de sistemas tecnológicos e na adaptação dos procedimentos internos à nova realidade da empresa, fez com que as Pequenas e Médias Empresas (PME) se mantivessem num circuito mais tradicional, ao contrário das grandes empresas. No entanto, o aumento da concorrência, a globalização dos mercados, o desenvolvimento tecnológico e a efemeridade das preferências dos clientes tem levado a que muitas PME repensem o seu negócio. Neste contexto, a questão de investigação definida foi: como é efectuada a adopção de CRM nas PME? Portanto, o objectivo foi: propor uma metodologia de implementação de CRM adaptada às necessidades das PME. Para que isso fosse possível a presente investigação seguiu o método de estudo de casos múltiplos, em duas empresas portuguesas. Como fonte de recolha de dados realizaram-se entrevistas e análise de documentos secundários. O principal contributo da tese é a proposta de uma metodologia de implementação de CRM capaz de responder às necessidades específicas das PME. Pelo facto de estas empresas deterem um peso importante no tecido empresarial Português e pelas poucas investigações realizadas neste âmbito...

Pectin methylesterase (PME), a ubiquitous enzyme in plants, de-esterifies the methoxylated pectin in the plant cell wall. We have characterized a PME gene (designated as pmeu1) from tomato (Lycopersicon esculentum) with an expression that is higher in younger root, leaf, and fruit tissues than in older tissues. Hypocotyls and epicotyls show higher accumulation of pmeu1 transcripts compared with cotyledons. pmeu1 represents a single-copy gene in the tomato genome. Comparison of the deduced amino acid sequence of pmeu1 with other PME homologs showed that the N-terminal halves are highly variable, and the C-terminal halves are relatively conserved in plant PMEs. Constitutive expression of a fruit-specific PME antisense gene does not affect the level of pmeu1 transcripts in vegetative tissues but does lower the level of PMEU1 mRNA in developing tomato fruits. These results suggest that there exists developmentally regulated silencing of pmeu1 by a heterologous PME antisense gene. Expression of pmeu1 in tobacco (Nicotiana tabacum) under the control of the cauliflower mosaic virus 35S promoter caused up to a 4-fold increase in PME specific activity that was correlated with the accumulation of PMEU1 mRNA. In vitro transcription-translation analyses show that pmeu1 encodes a 64-kD polypeptide...

The apical wall of growing pollen tubes must be strong enough to withstand the internal turgor pressure, but plastic enough to allow the incorporation of new membrane and cell wall material to support polarized tip growth. These essential rheological properties appear to be controlled by pectins, which constitute the principal component of the apical cell wall. Pectins are secreted as methylesters and subsequently deesterified by the enzyme pectin methylesterase (PME) in a process that exposes acidic residues. These carboxyls can be cross-linked by calcium, which structurally rigidifies the cell wall. Here, we examine the role of PME in cell elongation and the regulation of its secretion and enzymatic activity. Application of an exogenous PME induces thickening of the apical cell wall and inhibits pollen tube growth. Screening a Nicotiana tabacum pollen cDNA library yielded a pollen-specific PME, NtPPME1, containing a pre-region and a pro-region. Expression studies with green fluorescent protein fusion proteins show that the pro-region participates in the correct targeting of the mature PME. Results from in vitro growth analysis and immunolocalization studies using antipectin antibodies (JIM5 and JIM7) provide support for the idea that the pro-region acts as an intracellular inhibitor of PME activity...

BACKGROUND: In vivo hepatic phosphorus-31 magnetic resonance spectroscopy (31P MRS) provides biochemical information about phosphorus metabolism. AIM: To assess 31P MRS as a prognostic marker in patients with primary biliary cirrhosis (PBC) in relation to the current clinical prognostic models. PATIENTS AND METHODS: Twenty three patients with PBC of varying functional severity and 16 matched healthy volunteers were studied using in vivo 31P MRS. Spectra were acquired using a 1.5 T spectroscopy system. Peak area ratios of phosphomonoesters (PME), inorganic phosphate (Pi), and phosphodiesters (PDE) and nucleotide triphosphate (NTP) were calculated. Pugh score, Christensen prognostic index, and R value according to the Mayo model were calculated from the clinical data. RESULTS: The PME/NTP, Pi/NTP, PME/PDE, and PME/Pi ratios and the PME signal height ratio (SHR) were significantly higher, while the PDE/NTP and PDE/SHR were significantly lower in PBC patients compared with healthy volunteers (p < 0.01). Significant correlations were seen between PME/Pi ratio and the prognostic index according to Christensen (r = 0.63, p < 0.001), R value according to the Mayo model (r = 0.45, p < 0.03), and with the Pugh score (r = 0.55, p < 0.007). CONCLUSIONS: This study shows that PME/Pi ratio obtained from 31P MRS correlates well with all three of the commonly used models of prognosis in patients with PBC. A longitudinal study with larger number of patients is required to confirm these findings and elucidate the biochemical changes underlying this phenomenon.

Hepatic infiltration by lymphoma can be difficult to detect by conventional methods. We have studied 22 patients in vivo 31P magnetic resonance spectroscopy of the liver and compared the results with the clinical staging and assessment of liver involvement by computed tomography (CT), ultrasound (US), and liver function tests (LFTs). We find that the phosphomonoester (PME) to ATP, and the PME to Pi ratios are the best indication of liver involvement as in all the patients with liver involvement apparent on CT or US, these ratios were elevated (greater than 2 s.d. above the control mean). Of the patients with deranged LFTs but normal CT or US, five out of nine showed increased PME/ATP and PME/Pi ratios, and in the patients with normal LFTs and normal CT or US, three out of eight patients had raised PME ratios. Extracts of lymphomatous lymph nodes contain high concentrations of phosphoethanolamine which suggests that this compound is responsible for the increase in the PME peak. Eleven patients were studied again after chemotherapy, and those with initially raised PME/ATP and PME/Pi ratios all showed a decrease in these ratios towards normal. The patients with initially normal ratios showed no changes.

Patients infected with human immunodeficiency virus (HIV) often suffer from herpesvirus infections as a result of immunosuppression. These infections can occur while patients are receiving antiretroviral therapy, and additional drugs required to treat their infection can adversely affect compliance. It would be useful to have antivirals with a broader spectrum of activity that included both HIV and the herpesviruses. We reported previously that alkoxyalkyl ester prodrugs of cidofovir are up to 3 orders of magnitude more active against herpesvirus replication and may be less toxic than the unmodified drug. To determine if this strategy would be effective for certain phosphonomethoxyethyl nucleoside phosphonates which are also active against HIV infections, the hexadecyloxypropyl (HDP) esters of 1-(phosphonomethoxyethyl)-cytosine, 1-(phosphonomethoxyethyl)-5-bromo-cytosine (PME-5BrC), 1-(phosphonomethoxyethyl)-5-fluoro-cytosine, 9-(phosphonomethoxyethyl)-2,6-diaminopurine (PME-DAP), and 9-(phosphonomethoxyethyl)-2-amino-6-cyclopropylaminopurine (PME-cPrDAP) were evaluated for activity against herpesvirus replication. The HDP esters were substantially more active than the unmodified acyclic nucleoside phosphonates, indicating that esterification with alkoxyalkyl groups increases the antiviral activity of many acyclic nucleoside phosphonates. The most interesting compounds included HDP-PME-cPrDAP and HDP-PME-DAP...

Dietary intake of whole grains reduces the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. In an earlier study, we showed that Panicum miliaceum L. extract (PME) exhibited the highest anti-lipogenic activity in 3T3-L1 cells among extracts of nine different cereal grains tested. In this study, we hypothesized that PME in the diet would lead to weight loss and augmentation of hyperlipidemia by regulating fatty acid metabolism. PME was fed to ob/ob mice at 0%, 0.5%, or 1% (w/w) for 4 weeks. After the experimental period, body weight changes, blood serum and lipid profiles, hepatic fatty acid metabolism-related gene expression, and white adipose tissue (WAT) fatty acid composition were determined. We found that the 1% PME diet, but not the 0.5%, effectively decreased body weight, liver weight, and blood triglyceride and total cholesterol levels (P < 0.05) compared to obese ob/ob mice on a normal diet. Hepatic lipogenic-related gene (PPARα, L-FABP, FAS, and SCD1) expression decreased, whereas lipolysis-related gene (CPT1) expression increased in animals fed the 1% PME diet (P < 0.05). Long chain fatty acid content and the ratio of C18:1/C18:0 fatty acids decreased significantly in adipose tissue of animals fed the 1% PME diet (P < 0.05). Serum inflammatory mediators also decreased significantly in animals fed the 1% PME diet compared to those of the ob/ob control group (P < 0.05). These results suggest that PME is useful in the chemoprevention or treatment of obesity and obesity-related disorders.

To assess the coagulation capability of a perfusion microwave electrode (PME) as a key component of microwave coagulation therapy, a preliminary experimental study was performed using ex vivo and in vivo livers. For a microwave electrode, a PME was employed. Using a PME, saline was passed through the electrode and injected continuously into the target tissue. Using an ex vivo bovine liver, the range of tissue coagulation was measured for various volumes of infused saline and microwave outputs. Using an in vivo porcine liver, the efficiency of coagulation by a PME was compared with that of radiofrequency ablation (RFA) using a cool-tip needle. In an ex vivo bovine liver, the range of tissue coagulation increased as the flow rate of saline increased. In the in vivo porcine liver, the range of coagulation was similar to that found in the ex vivo bovine liver. With a PME under conditions of a microwave output of 80 W, a flow rate of 3 ml/min and irradiation time of 5 min, the range of coagulation was 44.8±2.8 mm [maximum vertical diameter: (a)] x 31.2±2.4 mm [maximum transverse diameter: (b)]. The range of RFA (cool-tip needle) at 12 min was 46.0±2.0 mm (a) x 30.2±2.0 mm (b). With only 5 min of microwave irradiation, the use of a PME enabled induction of the same range of coagulation that was obtainable by RFA for 12 min. In comparison with microwave coagulation without saline infusion...

Pectin is secreted in a highly methylesterified form and partially de-methylesterified in the cell wall by pectin methylesterases (PMEs). PME activity is expressed during plant growth, development and stress responses. PME activity is controlled at the post-transcriptional level by proteins named PME inhibitors (PMEIs). We have identified, expressed and characterized VvPMEI1, a functional PME inhibitor of Vitis vinifera. VvPMEI1 typically affects the activity of plant PMEs and is inactive against microbial PMEs. The kinetics of PMEI-PME interaction, studied by surface plasmon resonance, indicates that the inhibitor strongly interacts with PME at apoplastic pH while the stability of the complex is reduced by increasing the pH. The analysis of VvPMEI1 expression in different grapevine tissues and during grape fruit development suggests that this inhibitor controls PME activity mainly during the earlier phase of berry development. A proteomic analysis performed at this stage indicates a PME isoform as possible target of VvPMEI1.

Pectin methyl esterases (PMEs) and their endogenous inhibitors are involved in the regulation of many processes in plant physiology, ranging from tissue growth and fruit ripening to parasitic plant haustorial formation and host invasion. Thus, control of PME activity is critical for enhancing our understanding of plant physiological processes and regulation. Here we report on the identification of epigallocatechin gallate (EGCG), a green tea component, as a natural inhibitor for pectin methyl esterases. In a gel assay for PME activity, EGCG blocked esterase activity of pure PME as well as PME extracts from citrus and from parasitic plants. Fluorometric tests were used to determine the IC50 for a synthetic substrate. Molecular docking analysis of PME and EGCG suggests close interaction of EGCG with the catalytic cleft of PME . Inhibition of PME by the green tea compound, EGCG, provides means to study the diverse roles of PMEs in cell wall metabolism and plant development. In addition, this study introduces the use of EGCG as natural product to be used in the food industry and agriculture.; Other Research Unit

To examine tThe optimal conditions of cultivation for Cladosporium cladosporioides to produce and partially purify pectin methylesterase (PME) and polygalacturonase (PG) were examined under laboratory conditions . Activities of PME and PG were evaluated after 10, 15 and 20 days of incubation at 28°C in rice solid medium. Enzyme extraction was carried out by homogenization in buffer solutions at different pH (4, 5 and 6) followed by precipitation with ammonium sulfate [(NH4)2SO4] at 20, 40 and 60% saturation. After 10 days of incubation, the highest activities for PG (105.5 U/mL) and PME (1480 U/mL) in the crude extract was observed. Benzoate buffer (pH 4.0) was the best buffer for enzymes extraction. The final yield of 108.7% for PME and 10.6% for PG and a purification index of 14.2 and 1.4 for PME and PG, respectively, was obtained after saturation with 60%. The ideal conditions for obtained and pre-purification of PME and PG were 10 days of incubation, extraction with benzoate buffer pH 4.0 and precipitation with 60% of (NH4)2SO4. This is the first report on of pectinases pectinolytic enzymes produced by from C. cladosporioides and provides information about growth conditions for production and pre-purification of PME and PG.

Os solos podem funcionar como fonte ou sumidouro de carbono para a atmosfera, dependendo das condições climáticas e, sobretudo, do tipo de uso e manejo adotados. Levando-se em consideração o potencial em estocar/sequestrar C, reconhecidamente atribuídos a diversos usos da terra, como é o caso das florestas plantadas (Pinus e Eucaliptus), das áreas de plantio direto, bem como a eficiência em estocar C nos solos sob pastagens, é que se apresenta o objetivo da presente pesquisa: compreender a potencialidade em estocar C por diferentes usos da terra destacando: o reflorestamento (REF), a cultura anual irrigada (CAI); o plantio direto (PDI); o café (CAF), a cana-de-açúcar (CAN), a pastagem melhorada (PME) e a pastagem degradada (PDE), em comparação à vegetação natural de cerrado, mais especificamente a fitofisionomia denominada Cerrado/Cerradão (CER), que constitui a vegetação típica, no qual está inserida a bacia do rio Araguari-MG, para as profundidades 0-20 cm e 20-40 cm. A bacia do Rio Araguari, localizada na Mesorregião do Triângulo e Alto Paranaíba, porção oeste do Estado de Minas Gerais, está situada entre as coordenadas geográficas 18º 20’ e 20º 10’ de latitude Sul e 46º 00’ e 48º 50' de longitude Oeste de Greenwich...