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.

Variação no conteúdo de beta-glucanas em cultivares brasileiros de aveia

Com o crescente interesse em alimentos funcionais e nutracêuticos, a aveia (Avena sativa L.) tem se destacado, devido ao seu teor de fibras alimentares e principalmente às beta-glucanas. As (1,3)(1,4)-beta-D-glucanas, fibras alimentares na maioria solúveis, atuam na redução do colesterol em indivíduos com hipercolesterolemia. Existem estudos para determinar as causas de variação do teor desta fibra em aveia, porém, pouco se sabe sobre a aveia cultivada no Brasil. O objetivo deste trabalho foi verificar se existem diferenças no conteúdo de beta-glucanas entre cultivares brasileiros e se há variação na porcentagem desta fibra devido ao ano de cultivo. Os cultivares IAC7, UFRGS14, UPF16 e UPF17 (3 amostras de cada), e ainda três amostras do cultivar IAC7 para cada ano de cultivo (97 e 98), foram analisados segundo os métodos da AACC (American Association of Cereal Chemists). Os teores médios (peso seco) de beta-glucanas foram 6,50% (IAC7), 4,30% (UFRGS14), 3,51% (UPF16) e 3,78% (UPF17), com erro padrão de ±0,084 e coeficiente de variação de 7,89 %. Observou-se efeito significativo dos cultivares (p=0,03) e grande variabilidade entre as amostras (p=0,0001). O cultivar IAC7 apresentou média de beta-glucanas de 5,11% em 97 e 6,50 % em 98 (erro padrão ±0,14; CV=10,53%) e observou-se efeito significativo do ano de cultivo.

Alterações estruturais in vivo dos isômeros todo-trans, 9-cis e 13-cis do beta-caroteno

Com o objetivo de verificar alterações estruturais nos isômeros todo-trans, 9- e 13-cis do beta-caroteno foi realizado um ensaio biológico baseado no modelo de esgotamento das reservas hepáticas de carotenóides em ratos. Animais depletados desses carotenóides receberam, durante quinze dias, os isômeros puros todo-trans, 9-cis e 13-cis do beta-caroteno. Ao final deste período, verificou-se a ocorrência de re-isomerização in vivo desses isômeros, a partir da quantificação dos mesmos depositados no fígado dos animais. Foi observada re-isomerização do 9-cis em todo-trans, do todo-trans em 9-cis, do 13-cis em 9-cis e todo-trans. O 13-cis foi mais susceptível à isomerização que o 9-cis, pois este último passou a todo-trans e nunca a 13-cis. Já o 13-cis, tanto pode se transformar em 9-cis quanto em todo-trans.

Composição centesimal e teor de beta-glucanas em cereais e derivados

Foi utilizado o método enzimático recomendado pela AOAC para determinação de beta-glucanas em cereais e alimentos que os contém. O método, utiliza liquenase (EC 3.2.1.73) e beta-glucosidase (EC 3.2.1.21) para hidrólise debeta-glucanas, é rápido, fácil de executar e específico para beta-glucanas com ligações beta(1->3) e beta(1->4). As sementes analisadas foram subministradas pelo Instituto Agronômico de Campinas (IAC) e os alimentos adquiridos nos supermercados. Aveia e cevada são os grãos com maior conteúdo de beta-glucanas. Na aveia os teores determinados foram 6,48 e 5,94%. Nos 10 cultivares de cevada os teores de beta-glucanas oscilaram entre 2,04 e 9,68%. Trigo e triticale apresentaram teores de b-glucanas menores que 1%. Nos produtos comerciais o teor de beta-glucanas estava relacionado ao tipo de cereal da fórmula. O produto comercial de maior conteúdo de beta-glucanas é o farelo de aveia. As beta-glucanas são ingredientes funcionais em potencial e a conveniência ou não de estimular sua incorporação em alimentos deve ser mais estudada. Quanto à composição centesimal dos grãos de cereais, o teor de proteínas foi o que apresentou a maior variação e isso se reflete na composição dos produtos comerciais.

Biodisponibilidade do beta-caroteno da folha desidratada de mandioca (Manihot esculenta Crantz)

Com o objetivo de verificar a biodisponibilidade do beta-caroteno da folha de mandioca (Manihot esculenta Crantz) foi realizado um ensaio biológico baseado no modelo de esgotamento das reservas hepáticas de vitamina A em ratos. Um grupo de ratos depletados de vitamina A hepática recebeu folha desidratada de mandioca como fonte beta-caroteno durante 25 dias, e foram comparados com um grupo que recebeu ração com vitamina A, outro grupo com ração sem vitamina A e um último grupo com beta-caroteno. O grupo Zero foi constituído de 8 animais que receberam durante 15 dias ração à base de caseína, deficiente de vitamina A. Ao final do experimento todos os animais foram sacrificados e seus fígados e plasmas analisados por Cromatografia Líquida de Alta Eficiência (CLAE), sendo verificado que a biodisponibilidade do beta-caroteno da folha desidratada de mandioca foi baixa.

beta-caroteno, ácido ascórbico e antocianinas totais em polpa de frutos de aceroleira conservada por congelamento durante 12 meses

Objetivou-se avaliar as alterações de b-caroteno, ácido ascórbico e antocianinas totais na polpa de frutos de clones de aceroleira conservada por congelamento. Os frutos dos clones BRS 152 (Sertaneja) BRS 235 (Apodi), BRS 236 (Cereja), BRS 237 (Roxinha), BRS 238 (Frutacor) e II 47/1 foram colhidos no estádio de maturação comercial (vermelho maduro) em Limoeiro do Norte, Ceará, Brasil, transportados para a Planta Piloto de Processamento de Frutos da Embrapa Agroindústria Tropical, despolpados, acondicionada a polpa em sacos de polietileno (100 g), congelada, mantida em freezer a -20 °C, e avaliada a cada 30 dias durante 12 meses. O experimento foi realizado em delineamento experimental inteiramente casualizado em esquema fatorial 6 x 13 (clones x tempo), com 3 repetições. A concentração de beta-caroteno foi estável no clone Cereja, enquanto, nos demais, houve decréscimo durante todo o período do experimento. Houve pequeno decréscimo no teor de ácido ascórbico em todos os clones estudados durante o armazenamento, provavelmente devido à alta acidez da polpa, que auxilia na manutenção deste nutriente. O teor de antocianinas totais foi estável nos clones Frutacor e Sertaneja, enquanto nos demais houve diminuição. O clone II 47/1 foi dentre os estudados o que apresentou maiores teores de ácido ascórbico e antocianinas totais, mantendo estas características durante todo o armazenamento. De um modo geral, os clones em que se determinou menor teor de beta-caroteno foram observadas as mais elevadas concentrações de antocianinas totais.

Effect of processing on antioxidant potential and total phenolics content in beet (Beta vulgaris L.)

The antioxidant capacity of beet is associated with non-nutritive constituents, such as phenolic compounds. The purpose of this research was to evaluate the effect of two different heat-processing techniques (drying and canned) on the antioxidant potential (ABTS) and phenolics content of beets. A forced air circulation dehydrator was used for the drying. Drying at high temperatures (100 + 90 °C/5.6 hours; 90 °C/6 hours) increased the antioxidant potential of the processed products while mild drying conditions decreased it (80 °C/6 hours; 100 + 70 °C/6 hours) or had no effect on it (70 °C/7 hours; 100 + 80 °C/6 hours). For the canned products, the antioxidant potential did not differ according to the pH (4.2 to 3.8) for any of the four acids tested. Some processing methods influenced the antioxidant potential of the processed products, and this was also dependent on changes in the total phenolics content.

In-house method validation and occurrence of alpha-, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone and trifluralin residues in strawberry

A method for determination of organohalogen pesticides in strawberry by gas chromatography with electron capture detection was validated and applied in a monitoring program. Linearity, matrix effects, and day effect were evaluated for the analytes alpha-endosulfan, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone, and trifluralin. The linear range varied according to the chromatographic response of the analyte. Significant matrix effects were observed. The mean recoveries ranged from 74.6 to 115.4%, with repeatability standard deviations between 1.6 and 21.0% and intermediate precision between 5.9 and 21.0%. Detection, quantification and decision limit, and detection capacity ranged from 0.003 to 0.007 mg/kg, 0.005 to 0.013 mg/kg; 0.003 to 3.128 mg/kg; and 0.005 to 3.266 mg/kg, respectively. The method was fit for the purpose of monitoring organohalogen residues in strawberries. Residues of these pesticides were detected in 124 of the 186 samples analyzed between 2009 and 2011 in the state of Minas Gerais. Nine of them did not comply with the current legislation requirements; among them, seven (3.8%) had residues of unauthorized pesticide for the culture of strawberry, one (0.5%) had residues above the maximum residue limit, and another one (0.5%) exhibited both non-conformities.

Evaluation of beetroot (Beta vulgaris L.) leaves during its developmental stages: a chemical composition study

Beetroot leaves (Beta vulgaris L.) are commonly cut off and discarded before using its bulb due to lack of knowledge of how to use them. Aiming at using these leaves, in the present study, in natura and dehydrated beetroot leaves were chemically characterized in terms of fatty acid composition, proximate composition, minerals, total phenolic compounds (TPC), and antioxidant activity by DPPH• in different stages (60, 80, and 100 days) of development. The beetroot leaves showed significant levels of protein and lipids in all developmental stages, and all proximate composition nutrients decreased during these maturation stages; the highest content was observed at 60 days. The Fe content decreased during the developmental stages (from 342.75 to 246.30 mg.kg-1), while the content of K increased (from 13,367.64 to 20,784.90 mg.kg-1). With regard to to fatty acid composition, linolenic acid was present in the greatest quantity, and it increase up to 2.58 mg.g-1 (in natura) and 40.11 mg.g-1 (dehydrated) at 100 days of development. The n-6/n-3 ratios were low in all stages. The TPC and antioxidant activity by DPPH• changed during the developmental stages. The TPC was highest in the 100-day dehydrated leaves (15.27±0.12 mg GAE.g-1 FW), and the 50% inhibition of DPPH• (IC50 89.52 µg.mL-1) were better in the 60-day in natura leaves. This study shows that all developmental stages produced satisfactory results, and therefore, these leaves can be reused as food. The antioxidant activity and the chemical constituents, mainly the ω-3fatty acid, increased during the stages of development.

Alternative splicing of DNA polymerase beta in vertebrates

Alternative splicing (AS) is the predominant mechanism responsible for increasing eukaryotic transcriptome and proteome complexity. In this phenomenon, numerous mRNA transcripts are produced from a single pre-mRNA sequence. AS is reported to occur in 95% of human multi-exon genes; one specific gene that undergoes AS is DNA polymerase beta (POLB). POLB is the main DNA repair gene which performs short patch base excision repair (BER). In primate untransformed primary fibroblast cell lines, it was determined that the splice variant (SV) frequency of POLB correlates positively with species lifespan. To date, AS patterns of POLB have only been examined in mammals primarily through the use of cell lines. However, little attention has been devoted to investigating if such a relationship exists in non-mammals and whether cell lines reflect what is observed in vertebrate tissues. This idea was explored through cloning and characterization of 1,214 POLB transcripts from four non-mammalian species (Gallus gallus domesticus, Larus glaucescens, Xenopus laevis, and Pogona vitticeps) and two mammalian species (Sylvilagus floridanus and Homo sapiens) in two tissue types, liver and brain. POLB SV frequency occurred at low frequencies, < 3.2%, in non-mammalian tissues relative to mammalian (>20%). The highest POLB SV frequency was found in H. sapiens liver and brain tissues, occurring at 65.4% and 91.7%, respectively. Tissue specific AS of POLB was observed in L. glaucescens, P. vitticeps, and H. sapiens, but not G. gallus domesticus, X. laevis and S. floridanus.The AS patterns of a second gene, transient receptor potential cation channel subfamily V member 1 (TRPV1), were compared to those of POLB in liver and brain tissues of G. gallus domesticus, X. laevis and H. sapiens. This comparison was performed to investigate if any changes (either increase or decrease) observed in the AS of POLB were gene specific or if they were tissue specific, in which case similar changes in AS would be seen in POLB and TRPV1. Analysis did not reveal an increase or decrease in both the AS of POLB and TRPV1 in either the liver or brain tissues of G. gallus domesticus and H. sapiens. This result suggested that the AS patterns of POLB were not influenced by tissue specific rates of AS. Interestingly, an increase in the AS of both genes was only observed in X. laevis brain tissue. This result suggests that AS in general may be increased in the X. laevis brain as compared to liver tissue. No positive correlation between POLB SV frequency and species lifespan was found in non-mammalian tissues. The AS patterns of POLB in human primary untransformed fibroblast cell lines were representative of those seen in human liver tissue but not in brain tissue. Altogether, the AS patterns of POLB from vertebrate tissues and primate cell lines revealed a positive correlation between POLB SV frequency and lifespan in mammals, but not in non-mammals. It appears that this positive correlation does not exist in vertebrate species as a whole.