A role for protein kinase B beta/Akt2 in insulin-stimulated GLUT4 translocation in adipocytes

Insulin stimulates glucose uptake into muscle and fat cells by promoting the translocation of glucose transporter 4 (GLUT4) to the cell surface. Phosphatidylinositide 3-kinase (PI3K) has been implicated in this process. However, the involvement of protein kinase B (PKB)/Akt, a downstream target of PI3K in regulation of GLUT4 translocation, has been controversial. Here we report that microinjection of a PKB substrate peptide or an antibody to PKB inhibited insulin-stimulated GLUT4 translocation to the plasma membrane by 66 or 56%, respectively. We further examined the activation of PKB isoforms following treatment of cells with insulin or platelet-derived growth factor (PDGF) and found that PKB beta is preferentially expressed in both rat and 3T3-L1 adipocytes, whereas PKB alpha expression is down-regulated in 3T3-L1 adipocytes. A switch in growth factor response was also observed when 3T3-L1 fibroblasts were differentiated into adipocytes. While PDGF was more efficacious than insulin in stimulating PKB phosphorylation in fibroblasts, PDGF did not stimulate PKB beta phosphorylation to any significant extent in adipocytes, as assessed by several methods. Moreover, insulin, but not PDGF, stimulated the translocation of PKB beta to the plasma membrane and high-density microsome fractions of 3T3-L1 adipocytes. These results support a role for PKB beta in insulin-stimulated glucose transport in adipocytes.

PREDBALB/c: a system for the prediction of peptide binding to H2(d) molecules, a haplotype of the BALB/c mouse

PREDBALB/c is a computational system that predicts peptides binding to the major histocompatibility complex-2 (H2(d)) of the BALB/c mouse, an important laboratory model organism. The predictions include the complete set of H2(d) class I ( H2-K-d, H2-L-d and H2-D-d) and class II (I-E-d and I-A(d)) molecules. The prediction system utilizes quantitative matrices, which were rigorously validated using experimentally determined binders and non-binders and also by in vivo studies using viral proteins. The prediction performance of PREDBALB/c is of very high accuracy. To our knowledge, this is the first online server for the prediction of peptides binding to a complete set of major histocompatibility complex molecules in a model organism (H2(d) haplotype). PREDBALB/c is available at http://antigen.i2r.a-star.edu.sg/predBalbc/.

Mannan-binding lectin MBL2 gene polymorphism in chronic hepatitis C: association with the severity of liver fibrosis and response to interferon therapy

Hepatitis C virus (HCV) is a major cause of hepatic disease and of liver transplantation worldwide. Mannan-binding lectin (MBL), encoded by the MBL2 gene, can have an important role as an opsonin and complement activating molecule in HCV persistence and liver injury. We assessed the MBL2 polymorphism in 102 Euro-Brazilian patients with moderate and severe chronic hepatitis C, paired for gender and age with 102 HCV seronegative healthy individuals. Six common single nucleotide polymorphisms in the MBL2 gene, three in the promoter (H/L, X/Y and P/Q) and three in exon 1 (A, the wild-type, and B, C or D also known as O) were evaluated using real-time polymerase chain reaction with fluorescent hybridization probes. The concentration of MBL in plasma was measured by enzyme-linked immunosorbent assay. The frequency of the YA/YO genotype was significantly higher in the HCV patients compared with the controls (P = 0.022). On the other hand, the genotypes associated with low levels of MBL (XA/XA, XA/YO and YO/YO) were decreased significantly in the patients with severe fibrosis (stage F4), when compared with the patients with moderate fibrosis (stage F2) (P = 0.04) and to the control group (P = 0.011). Furthermore, MBL2 genotypes containing X or O mutations were found to be associated with non-responsiveness to pginterferon and ribavirin treatment (P = 0.023). MBL2 polymorphisms may therefore be associated not only with the development of chronic hepatitis C, but also with its clinical evolution and response to treatment.

Reciprocal interactions of obstructive sleep apnea and hypertension associated with ACE I/D polymorphism in males

Background: The angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism gene contributes to the genesis of hypertension (HTN) and may help explain the relationship between obstructive sleep apnea (OSA) and HTN. However, ACE is a pleiotropic gene that has several influences, including skeletal muscle and control of ventilation. We therefore tested the hypothesis that ACE polymorphism influences OSA severity. Methods: Male OSA patients (apnea-hypopnea index [AHI] > 5 events/h) from 2 university sleep centers were evaluated by polysomnography and ACE I/D polymorphism genotyping. Results: We studied 266 males with OSA (age = 48 +/- 13y, body mass index = 29 5kg/m(2), AHI = 34 +/- 25events/h). HTN was present in 114 patients (43%) who were older (p < 0.01), heavier (p < 0.05) and had more severe OSA (p < 0.01). The I allele was associated with HTN in patients with mild to moderate OSA (p < 0.01), but not in those with severe OSA. ACE I/D polymorphism was not associated with apnea severity among normotensive patients. In contrast. the only variables independently associated with OSA severity among patients with hypertension in multivariate analysis were BMI (OR = 1.12) and 11 genotype (OR = 0.27). Conclusions: Our results indicate reciprocal interactions between OSA and HTN with ACE I/D polymorphism, suggesting that among hypertensive OSA males, the homozygous ACE I allele protects from severe OSA. (C) 2009 Elsevier B.V. All rights reserved.

Quantitative ferromagnetic resonance analysis of CD133 stem cells labeled with iron oxide nanoparticles

The aim of this work is to provide a quantitative method for analysis of the concentration of superparamagnetic iron oxide nanoparticles (SPION), determined by means of ferromagnetic resonance (FMR), with the nanoparticles coupled to a specific antibody (AC133), and thus to express the antigenic labeling evidence for the stem cells C D133(+). The FMR efficiency and sensitivity were proven adequate for detecting and quantifying the low amounts of iron content in the C D133(+) cells (similar to 6.16 x 10(5) pg in the volume of 2 mu l containing 4.5 x 1011 SPION). The quantitative method led to the result of 1.70 x 10(-13) mol of Fe (9.5 pg), or 7.0 x 10(6) nanoparticles per cell. For the quantification analysis via the FMR technique it was necessary to carry out a preliminary quantitative visualization of iron oxide-labeled cells in order to ensure that the nanoparticles coupled to the antibodies are indeed tied to the antigen at the stem cell surface and that the cellular morphology was conserved, as proof of the validity of this method. The quantitative analysis by means of FMR is necessary for determining the signal intensity for the study of molecular imaging by means of magnetic resonance imaging (MRI).

Liver HLA-G expression is associated with multiple clinical and histopathological forms of chronic hepatitis B virus infection

As the mechanisms leading to the persistence of hepatitis B virus (HBV) infection are poorly understood and as the histocompatibility leucocyte antigen (HLA)-G is well described as a tolerogenic molecule, we evaluated HLA-G expression in 74 specimens of HBV liver biopsies and in 10 specimens obtained from previously healthy cadaver liver donors. HBV specimens were reviewed and classified by the METAVIR score, and HLA-G expression was assessed by immunohistochemistry. No HLA-G expression was observed in control hepatocytes. In contrast, 57 (77%) of 74 HBV specimens showed soluble and membrane-bound HLA-G expression in hepatocytes, biliary epithelial cells or both. No associations between the intensity of HLA-G expression and patient age or gender, HBeAg status, severity of liver fibrosis, and grade of histological findings were observed. Although significance was not reached (P = 0.180), patients exhibiting HLA-G expression presented a higher median HBV DNA viral load (105 copies/mL) than those who did not express HLA-G (103.7 copies/mL). These results indicate that HLA-G is expressed in most cases of chronic HBV infection in all stages and may play a role in the persistency of HBV infection.

The effect of ovariectomy on cardiac autonomic control in rats submitted to aerobic physical training

We have investigated the ovariectomy effects on the cardiovascular autonomic adaptations induced by aerobic physical training and the role played by nitric oxide (NO). Female Wistar rats (n =70) were divided into five groups: Sedentary Sham (SS): Trained Sham (TS); Trained Hypertensive Sham treated with N(C)-nitro-L-arginine methyl ester (L-NAME) (THS): Trained Ovariectomized (TO); and Trained Hypertensive Ovariectomized treated with L-NAME (THO). Trained groups were submitted to a physical training during 10 weeks. The cardiovascular autonomic control was investigated in all groups using different approaches: 1) pharmacological evaluation of autonomic tonus with methylatropine and propranolol; 2) analysis of heart rate (HR) and systolic arterial pressure (AP) variability; 3) spontaneous baroreflex sensitivity (BRS) evaluation. Hypertension was observed in THS and THO groups. Pharmacological analysis showed that TS group had increased predominance of autonomic vagal tonus compared to SS group. HR and intrinsic HR were found to be reduced in all trained animals. TS group, compared to other groups, showed a reduction in LF oscillations (LF=0.2-0.75 Hz) of pulse interval in both absolute and normalized units as well as an increase in HF oscillations (HF=0.75-2.50 Hz) in normalized unit. FIRS analysis showed that alpha-index was different between all groups. TS group presented the greatest value, followed by the TO, SS. THO and THS groups. Ovariectomy has negative effects on cardiac autonomic modulation in trained rats, which is characterized by an increase in the sympathetic autonomic modulation. These negative effects suggest NO deficiency. In contrast, the ovariectomy seems to have no effect on AP variability. (C) 2008 Elsevier B.V. All rights reserved.

GB virus C/Hepatitis G virus and other putative hepatitis non A-E viruses

The identification of the major agents causing human hepatitis (Hepatitis A, B, C, D and E Viruses) was achieved during the last 30 years. These viruses are responsible for the vast majority of human viral hepatitis cases, but there are still some cases epidemiologically related to infectious agents without any evidence of infection with known virus, designated as hepatitis non A - E. Those cases are considered to be associated with at least three different viruses: 1 - Hepatitis B Virus mutants expressing its surface antigen (HBsAg) with altered epitopes or in low quantities; 2 - Another virus probably associated with enteral transmitted non A-E hepatitis, called Hepatitis F Virus. Still more studies are necessary to better characterize this agent; 3 - Hepatitis G Virus or GB virus C, recently identified throughout the world (including Brazil) as a Flavivirus responsible for about 10% of parenteral transmitted hepatitis non A-E. Probably still other unknown viruses are responsible for human hepatitis cases without evidence of infection by any of these viruses, that could be called as non A-G hepatitis.

THE INFLUENCE OF HIV-1 SUBTYPES C, CRF31_BC AND B ON DISEASE PROGRESSION AND INITIAL VIROLOGIC RESPONSE TO HAART IN A SOUTHERN BRAZILIAN COHORT

Background: Although most HIV-1 infections in Brazil are due to subtype B, Southern Brazil has a high prevalence of subtype C and recombinant forms, such as CRF31_BC. This study assessed the impact of viral diversity on clinical progression in a cohort of newly diagnosed HIV-positive patients. Methods: From July/2004 to December/2005, 135 HIV-infected patients were recruited. The partial pol region was subtyped by phylogeny. A generalized estimating equation (GEE) model was used to examine the relationship between viral subtype, CD4+ T cell count and viral load levels before antiretroviral therapy. Hazard ratio (Cox regression) was used to evaluate factors associated with viral suppression (viral load < 50 copies/mL at six months). Results: Main HIV-1 subtypes included B (29.4%), C (28.2%), and CRF31_BC (23.5%). Subtypes B and C showed a similar trend in CD4+ T cell decline. Comparison of non-B (C and CRF31_BC) and B subtypes revealed no significant difference in the proportion of patients with viral suppression at six months (week 24). Higher CD4+ T cell count and lower viral load were independently associated with viral suppression. Conclusion: No significant differences were found between subtypes; however, lower viral load and higher CD4+ T cell count before therapy were associated with better response.

Desenvolvimento do produto X para doentes oncológicos

Em Portugal, são diagnosticados, anualmente, entre 40 a 45 mil novos casos de cancro. Esta incid��ncia é semelhante à que é observada nos restantes países da União Europeia e tem vindo a aumentar, nas últimas d��cadas. A prevalência de desnutrição em doentes oncológicos é muito elevada e está associada à diminuição da resposta aos tratamentos e qualidade de vida. O objectivo deste trabalho residiu na elaboração de um gelado hipercalórico e hiperproteico para doentes oncológicos, um gelado com uma composição nutricional adequada para melhorar ou manter o estado nutricional de doentes desnutridos ou em risco de desnutrição. Elaboraram-se quatro diferentes formulações de gelado de morango (A, B, C e D) que foram avaliadas, quantitativamente e qualitativamente, através de análise sensorial. Não se verificaram diferenças significativas entre os parâmetros avaliados para as quatro formulações, à excepção do parâmetro intensidade do frio. Apesar disso, através do cálculo do índice de aceitabilidade, as formulações dos gelados A e C foram as mais aceites pelos provadores. Posteriormente, procedeu-se à comparação, por análise sensorial, entre as formulações mais aceites e um suplemento líquido hipercalórico e hiperproteico de morango, já existente no mercado. Como resultado verificou-se que a maioria dos provadores considera os gelados como preferidos, para um maior número de parâmetros. A formulação de gelado C foi preferida por 70% dos provadores quando comparada com o suplemento líquido. Assim, conseguiu-se não só elaborar um gelado hipercalórico e hiperproteico, como também obter um produto com características que o tornam favorito, em relação a um produto muito semelhante e já existente no mercado.

Germinação de sementes de azedinha (Oxalis hirsutissima), espécie medicinal de Mato Grosso, Brasil

Sementes de azedinha (Oxalis hirsutissima Mart. & Zuc.) apresentam um baixo percentual de germinação, dificultando a propagação dessa espécie medicinal, tão utilizada pelas populações tradicionais da região contra as inflamações oculares. Sementes coletadas de uma população nativa em Poconé, no Estado de Mato Grosso, foram submetidas a dois ensaios de quebra de dormência. No primeiro foram usados 12 tratamentos: pré-embebição em água por 24, 36, 48, 69 e 96 horas, imersão em acetona por 10 e 40 minutos, imersão em ácido clorídrico por 10 e 40 segundos, imersão em álcool etílico 96° por 10 e 40 segundos, e sem pré-tratamento. No segundo ensaio foram usados 14 tratamentos: imersão em água quente (70 ºC) por 20, 40, 60 e 300 segundos; em água muito quente (85 ºC) por 20, 40, 120 e 300 segundos, em água fervendo (100 °C) por 10, 20,40, 120 e 300 segundos, e sem pré-tratamento. A imersão em ácido clorídrico por 40 segundos diferiu estatisticamente dos demais, e mesmo assim a porcentagem de germinação foi baixa (47%). A imersão em água quente (70 ºC) por 300 segundos e em água muito quente (85 ºC) por 40 segundos mostraram os melhores resultados, com 89 e 92% de germinação, respectivamente. A água fervendo (100 ºC) por 300 segundos reduziu a percentagem de germinação a 4%.

A concentração e a quantidade de micronutrientes e de alumínio no cafeeiro, Coffea arabica, L., variedade mundo novo (B.Rodr.) Choussy, aos dez anos de idade

O presente trabalho relata os dados obtidos sôbre a concentração e a quantidade de micronutrientes, boro (B), cloro (Cl), cobre (Cu), ferro (Fe), manganês (Mn), molibd��nio (Mo) e zinco (Zn), no tronco, ramos, fôlhas e frutos do cafeeiro, Coffea arabica, L., variedade mundo novo (B.Rodr.) Choussy, aos dez anos de idade e crescendo em solo latosólico da Escola Superior de Agricultura "Luiz de Queiroz", em Piracicaba. Além dos micronutrientes, são apresentados dados sobre a concentração e a quantidade de alumínio, nas citadas partes do cafeeiro.

Influência de cafés de gôsto rio em ligas com cafés de bebida mole

Os autores estudam a influência do café Rio em ligas com cafés brasileiros de bebida Mole. Foram ensaiadas porcentagens crescentes de Café Rio: 0,0; 0,5; 1,0; 1,5; 2,0; 2,5; 3,0; 3,5; 4,0; 4,5; 5,0; 7,5; 10,0; 12,5; 15,0; 20,0; 25,0; 30,0; 35,0; 40,0; 50,0. Realizaram-se dois experimentos em blocos incompletos equilibrados, com t = 21 tratamentos (os mencionados acima), k = 3 parcelas por bloco, r = 10 repetições, b = 70 blocos, L = 1. Cada parcela era formada de 3 xícaras, de tipo padrão, sobre as quais cada degustador dava uma só opinião. Cada parcela era provada por 3 degustadores. Os dados coletados são, pois, 630 para cada ensaio (210 parcelas, 3 degustadores). Atribuia-se a cada parcela, para fins de análise estatística, a média das opiniões dos 3 degustadores. Os dois ensaios deram resultados bem concordantes, que levaram às seguintes conclusões: a) Faz-se necessária a transformação dos dados, pois as variâncias relativas aos diversos tratamentos são muito discrepantes. b) A transformação T = √ Y d�� resultados satisfatórios. c) O café Rio prejudica sensivelmente a bebida do café Mole, para teores a partir de 2,0%. d) Para teores de 4,5% em diante a liga tem bebida Riada ou Rio. e) A regressão obtida não é estritamente linear, mas a linha reta da uma aproximação razoável. f) Consideradas as porcentagens de 0,0 a 10,0%,a equação de regressão para os dados transformados pela transformação T = √ Y é: T = 1,7045 - 0,127 X, onde X é a porcentagem de café Rio e T d�� a bebida, na escala numérica adotada, transformada pela raíz quadrada. g) A equação de regressão para os tratamentos de 0,0 a 10,0% de cafe Rio é Y = 3,0997 - 0,3281 X, isto é, há uma queda de 0,3281 na escala numérica da bebida, para cada unidade de porcentagem de café Rio.

Indicadors de pressió ambiental a les platges de Menorca: prova pilot (IPAPM'pp)

L’augment del turisme experimentat des de principis dels anys 70 a l’illa de Menorca (Illes Balears, Espanya) ha provocat un impacte sobre les seves platges, el seu principal atractiu turístic. Molts d���aquests impactes s’incrementen durant la temporada alta, compresa entre els mesos de maig a octubre. L’objectiu principal d���aquest projecte és l’elaboració d���un sistema d���indicadors de pressió ambiental útils per a l’anàlisi de les platges. La zona objecte d���estudi de la prova-pilot, correspon a les platges del sud-oest de Menorca, aquestes cales són les que actualment es troben sotmeses a una major pressió. Es tracta de catorze platges tipificades en tres categories (A, B, C) segons les característiques del tipus d���espai on es troben situades. Són les platges de: Cala Degollador, Cala Blanca, Cala’n Bosch, Son Xoriguer, Son Saura-Es Banyul, Son Saura-Bellavista, Es Talaier, Cala Turqueta, Cala Macarelleta, Cala Macarella, Cala Galdana, Cala Mitjana, Cala Trebalúger i Cala Escorxada. Partint d���un treball bibliogràfic, s’ha realitzat una selecció de mig centenar d���indicadors potencials dels quals catorze han format part dels IPAPM’pp, mitjançant l’elaboració d���una anàlisi multicriteri. Per a cada un dels IPAPM’pp s’ha desenvolupat una metodologia amb les corresponents fitxes individuals descriptives per al seu seguiment temporal. En aquest primer estudi, l’indicador que presenta un nombre de valors no acceptables en una major proporció de platges, és l’indicador 3. Índex de valoració de les mesures de conservació del sistema natural, seguit de l’indicador 6. Superfície subsistema sorra per usuari. El 2006, les platges de tipologia A, presenten un major percentatge positiu de les variables dels indicadors. Les platges de tipologia B i C presenten un percentatge menor d���acceptabilitat dels valors dels indicadors.