Glicerol como solvente verde em eficiente obtenção de beta-hidroxiésteres

In this work, we report a new method for obtaining racemic β-hydroxyesters by reduction of β-ketoesters. The use of glycerol as a reactional medium in selective reduction of β-ketoesters into the corresponding alcohols was shown to be a viable and more efficient alternative compared with the conventional methodology, taking into account green chemistry prerogatives.

Expressão do P16 e do PDGFR-Beta no adenocarcinoma gástrico

OBJETIVO: Detectar a expressão imunoistoquímica do p16 e do PDGFR-beta no adenocarcinoma gástrico. MÉTODOS: Foram estudados 36 pacientes submetidos a cirurgia para adenocarcinoma gástrico entre 1998 e 2002 no Hospital da Santa Casa de Porto Alegre. As variáveis investigadas foram: idade, sexo, tamanho e localização do tumor, número de linfonodos dissecados, número de linfonodos metastáticos, tipo histológico, extensão da ressecção cirúrgica e estadiamento patológico. RESULTADOS: Não foi detectada expressão do PDGFR-beta nas peças cirúrgicas. Em relação ao p16, detectou-se perda de expressão menor que 10% e menor que 1% respectivamente em 89% e 79% das peças estudadas. CONCLUSÃO: Não houve correlação entre a perda de p16 e as variáveis estudadas.

Relação entre a espessura endometrial e os níveis do beta-HCG com a resposta ao tratamento da gravidez ectópica com metotrexato

OBJETIVO: avaliar a relação entre os níveis séricos do hormônio gonadotrópico coriônico (beta-hCG) e a medida da espessura endometrial, em pacientes submetidas ao tratamento de gravidez ectópica com metotrexate. MÉTODOS: estudo clínico prospectivo, no qual foram avaliados os títulos do beta-hCG e a maior medida da espessura endometrial no eixo longitudinal uterino por meio da ultra-sonografia transvaginal, em 38 pacientes com gravidez ectópica íntegra, com diâmetro < 3,5 cm, com estabilidade hemodinâmica e aumento dos níveis do beta-hCG num intervalo de 24-48 horas. Todas receberam terapia com dose única de metotrexate (50 mg/m² intra muscular). Foram comparadas as médias dos valores do beta-hCG e da espessura endometrial, das mulheres que evoluíram com sucesso do tratamento e com aquelas que apresentaram insucesso terapêutico. Posteriormente, foi avaliada a diferença entre as médias dos valores séricos do beta-hCG segundo a espessura endometrial (<10,0 mm e >10,0 mm), independente da resposta terapêutica. Para análise estatística foi utilizado o teste t de Student. RESULTADOS: as médias dos valores iniciais do beta-hCG das 28 mulheres que apresentaram resposta terapêutica medicamentosa foi de 1936,2 mUI/ml, significativamente menor do que a média das 10 mulheres sem resposta, 6831,3 mUI/ml (<0,04). Da mesma forma a média da espessura do endométrio foi significativamente menor nas mulheres com resposta d que naquelas sem resposta, 6,4 mm e 11,7mm respectivamente (p<0,01). A média dos valores do beta-hCG das mulheres com espessura endometrial <10,0 mm foi 2008,7 mUI/ml, significativamente menor que daquelas com endométrio >10,0 mm, cuja média foi de 6925,9 mUI/ml. CONCLUSÕES: a medida da espessura endometrial avaliada pela ultra-sonografia sofre influência dos níveis séricos do beta-hCG, se mostrando valiosa como fator orientador adicional na indicação do tratamento medicamentoso com metotrexate na gravidez ectópica íntegra.

Avaliação da função das células beta pancreáticas através do modelo matemático de HOMA em portadoras de síndrome dos ovários policísticos: comparação entre obesas e não-obesas

OBJETIVO: avaliar o efeito da obesidade sobre a função das célulasbeta pancreáticas de pacientes portadoras de síndrome dos ovários policísticos (SOP). MÉTODOS: estudo transversal no qual foram avaliadas 82 pacientes portadoras de SOP, selecionadas de forma consecutiva, no momento do diagnóstico de SOP. Pacientes com índice de massa corporal (IMC) maior ou igual a 30 kg/m² foram consideradas SOP obesas (n=31). Valores de índice de massa corporal menores que este limite foram consideradas SOP não-obesas, o que correspondeu a 51 mulheres. Foram utilizadas a glicemia e a insulina de jejum para cálculo da função das células beta pancreáticas (HOMA-%beta-Cell) e da resistência à insulina (HOMA-IR e QUICKI) entre os grupos. Analisaram-se, também, variáveis secundárias como idade, idade da menarca, níveis séricos hormonais (testosterona, prolactina, LH e FSH) e de colesterol total, triglicerídeos, HDL colesterol e LDL-colesterol. RESULTADOS: a idade da menarca das pacientes obesas com SOP (11,7±1,8 anos) foi menor que as não-obesas (12,7±1,9) (p<0,05). As SOP obesas tiveram LH inferior (7,9±5,0 mUI/mL) ao valor encontrado nas não-obesas (10,6±6,0 mUI/mL) (p<0,05). Ambos os grupos apresentaram a média de HDL colesterol inferior a 50 mg/dL. As pacientes obesas apresentaram insulina basal (32,5±25,2 mUI/mL) e glicemia de jejum (115,9±40,7 mg/dL) mais elevadas que as não-obesas (8,8±6,6 mUI/mL e 90,2±8,9 mg/dL, respectivamente) (p<0,01). No grupo SOP obesas, a freqüência de resistência à insulina foi de 93 versus 25% no grupo SOP não-obesas (p<0,01). Foi verificada hiperfunção das células beta do pâncreas em 86% das obesas com SOP contra 41% das não-obesas portadoras de SOP (p<0,0001). CONCLUSÕES: as pacientes com SOP obesas apresentaram freqüência mais alta de resistência à insulina e hiperfunção de células beta do pâncreas quando comparadas com pacientes SOP não-obesas.

Lead (Pb2+) and cadmium (Cd2+) inhibit the dipsogenic action of central beta-adrenergic stimulation by isoproterenol

We have previously demonstrated that acute third ventricle injections of both Pb2+ and Cd2+ impair the dipsogenic response elicited by three different situations: dehydration and central cholinergic or angiotensinergic stimulation. ß-Adrenergic activation is part of the multifactorial integrated systems operating in drinking behavior control in the central nervous system. In the present study acute third ventricle injections of Pb2+ (3, 30 and 300 pmol/rat) or Cd2+ (0.3, 3 and 30 pmol/rat) blocked the dipsogenic response induced by third ventricle injections of isoproterenol (ISO; 160 nmol/rat) in a dose-dependent manner. Normohydrated animals receiving ISO + NaAc (sodium acetate) or saline (controls) displayed a high water intake after 120 min (ISO + saline = 5.78 ± 0.54 ml/100 g; ISO + NaAc = 6.00 ± 0.6 ml/100 g). After the same period, animals receiving ISO but pretreated with PbAc at the highest dose employed (300 pmol/rat) drank 0.78 ± 0.23 ml/100 g while those receiving ISO and pretreated with the highest dose of CdCl2 (30 pmol/rat) presented a water intake of 0.7 ± 0.30 ml/100 g. Third ventricle injections of CdCl2 (3 nmol/rat) or PbAc (3 nmol/rat) did not modify food intake in rats deprived of food for 24 h. Thus, general central nervous system depression explaining the antidipsogenic action of the metals can be safely excluded. It is concluded that both Pb2+ and Cd2+ inhibit water intake induced by central ß-adrenergic stimulation

Metabolic markers following beta-adrenoceptor agonist infusion in footshock-stressed rats

Stress hormones can alter metabolic functions in adipose tissue and liver, as well as the sensitivity of rat white adipocytes and rat atrial responses to ß-adrenergic agonists. In this study, we examined the effects of three daily footshock stress sessions on the plasma corticosterone, glucose, glycerol and triacylglycerol levels of fed, conscious male rats, and on the plasma glucose, glycerol and triacylglycerol levels of the same rats following iv infusions of ß-adrenergic agonists (isoproterenol: 0.4 nmol kg-1 min-1, noradrenaline: 5.0 µg kg-1 day-1, and BRL 37344 ([±]-[4-(2-[(2-[3-chlorophenyl]-2-hydroxyethyl)amino]propyl)phenoxy]acetic acid), a selective ß3-adrenoceptor agonist: 0.4 nmol kg-1 min-1). Plasma corticosterone levels increased significantly after each stress session, while triacylglycerol levels increased after the first session and glucose increased after the second and third sessions. Glycerol levels were unaltered after stress. These results suggest that repeated footshock stress may induce a metabolic shift from triacylglycerol biosynthesis to glucose release by hepatic tissue, with glycerol serving as one of the substrates in both pathways. Stressed rats were more sensitive to infusion of noradrenaline plus prazosin and to infusion of isoproterenol, with elevated plasma glucose, glycerol and triacylglycerol levels. The higher sensitivity of stressed rats to isoproterenol and noradrenaline was probably related to the permissive effect of plasma corticosterone. Only BRL 37344 increased plasma glycerol levels in stressed rats, probably because ß3-adrenoceptors are not involved in hepatic triacylglycerol synthesis, thus allowing glycerol to accumulate in plasma.

Development of a Glycocluster Adjuvant Mimicking Natural Yeast beta-(1,2)-Structures

Allergy is characterized by T helper (Th) 2-type immune response after encounter with an allergen leading to subsequent immunoglobulin (Ig) E-mediated hypersensitivity reaction and further allergic inflammation. Allergen-specific immunotherapy (SIT) balances the Th2-biased immunity towards Th1 and T regulatory responses. Adjuvants are used in allergen preparations to intensify and modify SIT. β-(1,2)-oligomannoside constituents present in Candida albicans (C. albicans) cell wall possess Th1-type immunostimulatory properties. The aim of this thesis was to develop a β-(1,2)-linked carbohydrate compound with known structure and anti-allergic properties to be applied as an adjuvant in SIT. First the immunostimulatory properties of various fungal extracts were studied. C. albicans appeared to be the most promising Th1-inducing extract, which led to the synthesis of various mono- or divalent oligomannosides designed on the basis of C. albicans. These carbohydrates did not induce strong cytokine production in human peripheral blood mononuclear cell (PBMC) cultures. In contrast to earlier reports using native oligosaccharides from C. albicans, synthetic -(1,2)-linked mannotetraose did not induce any tumor necrosis factor production in murine macrophages. Next, similarities with synthesized divalent mannosides and the antigenic epitopes of β-(1,2)-linked C. albicans mannan were investigated. Two divalent compounds inhibited specific IgG antibodies binding to below 3 kDa hydrolyzed mannan down to the level of 30–50% showing similar antigenicity to C. albicans. Immunomodulatory properties of synthesized carbohydrate assemblies ranging from mono- to pentavalent were evaluated. A trivalent acetylated dimannose (TADM) induced interleukin-10 (IL-10) and interferon-γ responses. TADM also suppressed birch pollen induced IL-4 and IL-5 responses in allergen (Bet v) stimulated PBMCs of birch pollen allergic subjects. This suppression was stronger with TADM than with other used adjuvants, immunostimulatory oligonucleotides and monophosphoryl lipid A. In a murine model of asthma, the allergen induced inflammatory responses could also be suppressed by TADM on cytokine and antibody levels.

Impaired beta-adrenergic response and decreased L-type calcium current of hypertrophied left ventricular myocytes in postinfarction heart failure

Infarct-induced heart failure is usually associated with cardiac hypertrophy and decreased ß-adrenergic responsiveness. However, conflicting results have been reported concerning the density of L-type calcium current (I Ca(L)), and the mechanisms underlying the decreased ß-adrenergic inotropic response. We determined I Ca(L) density, cytoplasmic calcium ([Ca2+]i) transients, and the effects of ß-adrenergic stimulation (isoproterenol) in a model of postinfarction heart failure in rats. Left ventricular myocytes were obtained by enzymatic digestion 8-10 weeks after infarction. Electrophysiological recordings were obtained using the patch-clamp technique. [Ca2+]i transients were investigated via fura-2 fluorescence. ß-Adrenergic receptor density was determined by [³H]-dihydroalprenolol binding to left ventricle homogenates. Postinfarction myocytes showed a significant 25% reduction in mean I Ca(L) density (5.7 ± 0.28 vs 7.6 ± 0.32 pA/pF) and a 19% reduction in mean peak [Ca2+]i transients (0.13 ± 0.007 vs 0.16 ± 0.009) compared to sham myocytes. The isoproterenol-stimulated increase in I Ca(L) was significantly smaller in postinfarction myocytes (Emax: 63.6 ± 4.3 vs 123.3 ± 0.9% in sham myocytes), but EC50 was not altered. The isoproterenol-stimulated peak amplitude of [Ca2+]i transients was also blunted in postinfarction myocytes. Adenylate cyclase activation through forskolin produced similar I Ca(L) increases in both groups. ß-Adrenergic receptor density was significantly reduced in homogenates from infarcted hearts (Bmax: 93.89 ± 20.22 vs 271.5 ± 31.43 fmol/mg protein in sham myocytes), while Kd values were similar. We conclude that postinfarction myocytes from large infarcts display reduced I Ca(L) density and peak [Ca2+]i transients. The response to ß-adrenergic stimulation was also reduced and was probably related to ß-adrenergic receptor down-regulation and not to changes in adenylate cyclase activity.

Simultaneous changes in the function and expression of beta 1 integrins during the growth arrest of poorly differentiated colorectal cells (LISP-1)

Cells usually lose adhesion and increase proliferation and migration during malignant transformation. Here, we studied how proliferation can affect the other two characteristics, which ultimately lead to invasion and metastasis. We determined the expression of ß1 integrins, as well as adhesion and migration towards laminin-1, fibronectin, collagens type I and type IV presented by LISP-1 colorectal cancer cells exposed to 2.5% dimethyl sulfoxide (DMSO), an agent capable of decreasing proliferation in this poorly differentiated colorectal cell line. Untreated cells (control), as shown by flow cytometry and monoclonal antibodies, expressed alpha2 (63.8 ± 11.3% positive cells), alpha3 (93.3 ± 7.0%), alpha5 (50.4 ± 12.0%) and alpha6 (34.1 ± 4.9%) integrins but not alpha1, alpha4, alphav or ß4. Cells adhered well to laminin-1 (73.4 ± 6.0%) and fibronectin (40.0 ± 2.0%) substrates but very little to collagens. By using blocking monoclonal antibodies, we showed that alpha2, alpha3 and alpha6 mediated laminin-1 adhesion, but neither alpha3 nor alpha5 contributed to fibronectin adherence. DMSO arrested cells at G0/G1 (control: 55.0 ± 2.4% vs DMSO: 70.7 ± 2.5%) while simultaneously reducing alpha5 (24.2 ± 19%) and alpha6 (14.3 ± 10.8%) expression as well as c-myc mRNA (7-fold), the latter shown by Northern blotting. Although the adhesion rate did not change after exposure to DMSO, alpha3 and alpha5 played a major role in laminin-1 and fibronectin adhesion, respectively. Migration towards laminin-1, which was clearly increased upon exposure to DMSO (control: 6 ± 2 cells vs DMSO: 64 ± 6 cells), was blocked by an antibody against alpha6. We conclude that the effects of DMSO on LISP-1 proliferation were accompanied by concurrent changes in the expression and function of integrins, consequently modulating adhesion/migration, and revealing a complex interplay between function/expression and the proliferative state of cells.

Non-obese adult onset diabetes with oral hypoglycemic agent failure: islet cell autoantibodies or reversible beta cell refractoriness?

Pancreatic ß cell function and insulin sensitivity, analyzed by the homeostasis model assessment, before and after 24 weeks of insulin therapy were studied and correlated with the presence of autoantibodies against ß cells (islet cell and anti-glutamic acid decarboxylase antibodies), in a group of 18 Brazilian lean adult non-insulin-dependent diabetes mellitus (NIDDM) patients with oral hypoglycemic agent failure (OHAF). Median fasting plasma glucose before and after insulin treatment was 19.1 and 8.5 mmol/l, respectively (P < 0.001); median HbA1c was 11.7% before vs 7.2% after insulin treatment (P < 0.001). Forty-four percent of the patients were positive (Ab+) to at least one autoantibody. Fasting C-peptide levels were lower in Ab+ than Ab- patients, both before (Ab+: 0.16 ± 0.09 vs Ab-: 0.41 ± 0.35 nmol/l, P < 0.003) and after insulin treatment (Ab+: 0.22 ± 0.13 vs Ab-: 0.44 ± 0.24 nmol/l, P < 0.03). Improvement of Hß was seen in Ab- (median before: 7.3 vs after insulin therapy: 33.4%, P = 0.003) but not in Ab+ patients (median before: 6.6 vs after insulin therapy: 20.9%). These results show that the OHAF observed in the 18 NIDDM patients studied was due mainly to two major causes: autoantibodies and ß cell desensitization. Autoantibodies against ß cells could account for 44% of OHAF, but Ab- patients may still present ß cell function recovery, mainly after a period of ß cell rest with insulin therapy. However, the effects of ß cell function recovery on the restoration of the response to oral hypoglycemic agents need to be determined.

Antifungal activity of the naphthoquinone beta-lapachone against disseminated infection with Cryptococcus neoformans var. neoformans in dexamethasone-immunosuppressed Swiss mice

The in vivo antifungal activity of the naphthoquinone beta-lapachone against disseminated infection by Cryptococcus neoformans was investigated. Swiss mice were immunosuppressed daily with dexamethasone (0.5 mg per mouse) intraperitoneally for 3 days, the procedure was repeated 4 days later, and the animals were then challenged intravenously with C. neoformans (10(6) CFU/mL) 1 week later. Seven days after infection, the mice were divided into groups and treated daily with beta-lapachone (10 mg/kg, iv) for 7 (N = 6) and 14 days (N = 10). Amphotericin B (0.5 mg/kg) was used as comparator drug and an additional group received PBS. Treatment with beta-lapachone cleared the yeast from the spleen and liver, and the fungal burden decreased approximately 10(4) times in the lungs and brain 14 days after infection when compared to the PBS group (P < 0.05). This result was similar to that of the amphotericin B-treated group. Protection was suggestively due to in vivo antifungal activity of this drug and apparently not influenced by activation of the immune response, due to similar leukocyte cell counts among all groups. This study highlights the prospective use of beta-lapachone for treatment of disseminated cryptococcosis.

Short-term follow-up of exercise training program and beta-blocker treatment on quality of life in dogs with naturally acquired chronic mitral valve disease

This study aimed to evaluate the effects of carvedilol treatment and a regimen of supervised aerobic exercise training on quality of life and other clinical, echocardiographic, and biochemical variables in a group of client-owned dogs with chronic mitral valve disease (CMVD). Ten healthy dogs (control) and 36 CMVD dogs were studied, with the latter group divided into 3 subgroups. In addition to conventional treatment (benazepril, 0.3-0.5 mg/kg once a day, and digoxin, 0.0055 mg/kg twice daily), 13 dogs received exercise training (subgroup I; 10.3±2.1 years), 10 dogs received carvedilol (0.3 mg/kg twice daily) and exercise training (subgroup II; 10.8±1.7 years), and 13 dogs received only carvedilol (subgroup III; 10.9±2.1 years). All drugs were administered orally. Clinical, laboratory, and Doppler echocardiographic variables were evaluated at baseline and after 3 and 6 months. Exercise training was conducted from months 3-6. The mean speed rate during training increased for both subgroups I and II (ANOVA, P>0.001), indicating improvement in physical conditioning at the end of the exercise period. Quality of life and functional class was improved for all subgroups at the end of the study. The N-terminal pro-brain natriuretic peptide (NT-proBNP) level increased in subgroup I from baseline to 3 months, but remained stable after training introduction (from 3 to 6 months). For subgroups II and III, NT-proBNP levels remained stable during the entire study. No difference was observed for the other variables between the three evaluation periods. The combination of carvedilol or exercise training with conventional treatment in CMVD dogs led to improvements in quality of life and functional class. Therefore, light walking in CMVD dogs must be encouraged.

Purificação de três diferentes beta-galactosidades microbianas por partição em sistemas de duas fases aquosas

Este trabalho tratou da investigação do efeito do peso molecular de polietilenoglicol (PEG) sobre a partição de enzimas beta-galactosidases de diferentes origens microbianas: Escherichia coli, Klueveromyces lactis e Aspergillus orizae em sistemas de duas fases aquosas (SDFA).Foi observado que os melhores sistemas para purificação da enzima de E. coli foram os formados por PEG 4000, 6000 e 8000/fosfato, fornecendo os mais elevados fatores de purificação da enzima. As enzimas de Klueveromyces lactis e Aspergillus orizae não foram eficientemente purificadas nestes sistemas sendo insensíveis à alterações do peso molecular do PEG. Portanto, um outro sistema de duas fases aquosas foi desenvolvido contendo um ligante específico, p-aminofenil 1-tio-beta-D-galactopiranosídeo (APGP), acoplado ao PEG para purificar a enzima de Klueveromyces lactis. Uma etapa simples de partição no SDFA formado por 6% APGP-PEG4000 + 12% dextrana T505.000 foi capaz de recuperar 83% da enzima na fase superior do sistema e de aumentar 1,6 vezes o fator de purificação.

Teores de retinol, beta-caroteno e alfa-tocoferol em leites bovinos comercializados na cidade de São Paulo

Os teores de retinol, beta-caroteno e alfa-tocoferol foram determinados por cromatografia líquida de alta eficiência em leites em pó, pasteurizados e esterilizados, comercializados na Cidade de São Paulo. Após a saponificação e extração, os compostos foram determinados simultaneamente utilizando-se coluna de sílica, fase móvel constituída por hexano:isopropanol (99:1) e fluxo de 2,0mL/min. O retinol e o beta-caroteno foram determinados no detector UV/visível e o alfa-tocoferol no detector de fluorescência, ligado em série com o anterior. Os valores de vitamina A dos leites foram calculados com e sem a consideração do beta-caroteno. A maior contribuição deste nutriente no valor de vitamina A esteve entre os leites em pó, cerca de 17% em uma das marcas. Os altos teores das vitamina A e E encontrados em alguns leites, indicam que os mesmos provavelmente receberam adição destas vitaminas, não trazendo, entretanto, tal informação no rótulo. A análise de vitaminas nestes produtos indica a necessidade de maior controle de qualidade dos mesmos.

Avaliação dos teores de fibra alimentar e de beta-glicanas em cultivares de aveia (Avena sativa L)

A fibra alimentar é composta por celulose, hemiceluloses, gomas, pectinas e mucilagens sendo classificada em solúvel e insolúvel, quanto a sua solubilidade em água. As beta-glicanas são componentes da fibra alimentar solúvel presentes na aveia e sua importância é devido às propriedades funcionais e aos efeitos hipocolesterolêmicos e hipoglicêmicos apresentados. O presente trabalho tem como objetivo avaliar os teores de fibra alimentar solúvel, insolúvel e total e de beta-glicanas de cultivares de aveia recomendados pela Comissão Brasileira de Pesquisa de Aveia. Grãos de aveia (Avena sativa, L) foram descascados, as cariopses moídas e as amostras acondicionadas e armazenadas à temperatura de -20° C. Para a análise de fibra alimentar foi adotada a metodologia da AOAC (1997). Entre os cultivares analisados, UPF 7, UPF 13, UPF 14 e UPF 16 apresentaram os maiores teores de fibra alimentar insolúvel. Os maiores teores de fibra alimentar solúvel foram verificados nos cultivares UFRGS 7, CTC 13, UPF 16 e CTC 2. O cultivar UPF 16 apresentou o maior teor de fibra alimentar total, seguido de UFRGS 7, CTC 13 e UFRGS 18. Para a determinação de beta-glicanas foi adotada a metodologia da AOAC (1997). Os maiores teores de beta-glicanas foram verificados nos cultivares UFRGS 7, UPF 14 e UFRGS 18.