Serum amyloid A is a growth factor for 3T3-L1 adipocytes, inhibits differentiation and promotes insulin resistance

BACKGROUND/OBJECTIVES: Serum amyloid A (SAA) is an acute-phase protein that has been recently correlated with obesity and insulin resistance. Therefore, we first examined whether human recombinant SAA (rSAA) could affect the proliferation, differentiation and metabolism of 3T3-L1 preadipocytes. DESIGN: Preadipocytes were treated with rSAA and analyzed for changes in viability and [H-3-methyl]-thymidine incorporation as well as cell cycle perturbations using flow cytometry analysis. The mRNA expression profiles of adipogenic factors during the differentiation protocol were also analyzed using real-time PCR. After differentiation, 2-deoxy-[1,2-H-3]-glucose uptake and glycerol release were evaluated. RESULTS: rSAA treatment caused a 2.6-fold increase in cell proliferation, which was consistent with the results from flow cytometry showing that rSAA treatment augmented the percentage of cells in the S phase (60.9 +/- 0.54%) compared with the control cells (39.8 +/- 2.2%, ***P<0.001). The rSAA-induced cell proliferation was mediated by the ERK1/2 signaling pathway, which was assessed by pretreatment with the inhibitor PD98059. However, the exposure of 3T3-L1 cells to rSAA during the differentiation process resulted in attenuated adipogenesis and decreased expression of adipogenesis-related factors. During the first 72 h of differentiation, rSAA inhibited the differentiation process by altering the mRNA expression kinetics of adipogenic transcription factors and proteins, such as PPAR gamma 2 (peroxisome proliferator-activated receptor gamma 2), C/EBP beta (CCAAT/enhancer-binding protein beta) and GLUT4. rSAA prevented the intracellular accumulation of lipids and, in fully differentiated cells, increased lipolysis and prevented 2-deoxy-[1,2-H-3]-glucose uptake, which favors insulin resistance. Additionally, rSAA stimulated the secretion of proinflammatory cytokines interleukin 6 and tumor necrosis factor alpha, and upregulated SAA3 mRNA expression during adipogenesis. CONCLUSIONS: We showed that rSAA enhanced proliferation and inhibited differentiation in 3T3-L1 preadipocytes and altered insulin sensitivity in differentiated cells. These results highlight the complex role of SAA in the adipogenic process and support a direct link between obesity and its co-morbidities such as type II diabetes.

SUPERAÇÃO DE DORMÊNCIA DAS SEMENTES E CONTROLE QUÍMICO DE Momordicacharantia L.

Objetivou-se com este estudo avaliar diferentes métodos para a superação da dormência das sementes e a eficiência de herbicidas no controle químico de plantas de Momordicacharantia L. no estádio reprodutivo. O primeiro experimento constituiu-seem esquema fatorial 2 x 8 (duas épocas de coleta das sementes (2006 e 2007) e oito métodos de quebra de dormência: escarificação mecânica; imersão em ácido sulfúrico concentrado e a 50%; imersão em nitrato de potássio a 2%por três e seis horas; calor seco a 60°C; água quente a 60°C e uma testemunha sem tratamento). No segundo ensaio avaliou-se o controle da Momordicacharantia L. por meio de seis herbicidas distintos: Imazapic, Metsulfuron-methyl, Metribuzin, 2,4-D, Amicarbazone, Paraquat e uma testemunha sem aplicação de herbicidas. Em ambos os ensaios, o delineamento experimental utilizado foi o inteiramente casualizados, com quatro repetições. Após a aplicação dos tratamentos,pode-se concluir que a imersão das sementes em ácido sulfúrico concentrado por 3 minutos proporcionou uma maior porcentagem e velocidade de germinação e que o herbicida Metribuzin alcançou, embora tardiamente, um controle total das plantas de melão de São-Caetano.

Efeito de restritores hídricos sobre a germinação, comprimento da radícula e níveis de detecção de Colletotrichum gossypii var. cephalosporioides em sementes de algodão

O fungo Colletotrichum gossypii var. cephalosporioides, agente causal da ramulose do algodoeiro, é transmitido pela semente que se constitui em uma das mais importantes fontes de inóculo inicial e de introdução da doença em áreas indenes. Para que se possa identificar sua presença em lotes de sementes, é importante que se empreguem métodos de detecção rápidos e seguros. O mais empregado é o do papel de filtro, que se baseia na avaliação de sinais do patógeno desenvolvidos sobre as sementes, seguida da sua identificação morfológica. O método apresenta a desvantagem do crescimento das plântulas no período de incubação das sementes que pode favorecer o desenvolvimento de outros fungos e prejudicar a caracterização do patógeno. Para minimizar este problema vem sendo empregada a técnica da restrição hídrica. O presente trabalho teve como objetivo avaliar o efeito de três solutos em dois potenciais osmóticos, comparados ao tratamento padrão de água destilada, ao congelamento e ao 2,4 D, sobre a germinação, comprimento da radícula e detecção do agente causal da ramulose, durante o teste de sanidade. Os solutos Manitol e NaCl foram mais eficientes em inibir a germinação e favorecer a incidência do patógeno no potencial osmótico de -0,8 MPa. O KCl mostrou-se eficiente em inibir a germinação nos dois potenciais osmóticos testados, -0,6 e -0,8 MPa, porém reduziu a incidência do patógeno no potencial de -0,8 MPa. Os solutos Manitol, nos potenciais osmóticos de -0,8 e -0,6 MPa e o NaCl no potencial osmótico de -0,8 foram eficientes em reduzir o comprimento da radícula, sem interferir negativamente nos níveis de detecção de C. gossypii var. cephalosporioides, podendo ser recomendados para uso em análises sanitárias de rotina.

Cessation of physical exercise changes metabolism and modifies the adipocyte cellularity of the periepididymal white adipose tissue in rats

All of the adaptations acquired through physical training are reversible with inactivity. Although significant reductions in maximal oxygen uptake (VO2max) can be observed within 2 to 4 wk of detraining, the consequences of detraining on the physiology of adipose tissue are poorly known. Our aim was therefore to investigate the effects of discontinuing training (physical detraining) on the metabolism and adipocyte cellularity of rat periepididymal (PE) adipose tissue. Male Wistar rats, aged 6 wk, were divided into three groups and studied for 12 wk under the following conditions: 1) trained (T) throughout the period; 2) detrained (D), trained during the first 8 wk and detrained during the remaining 4 wk; and 3) age-matched sedentary (S). Training consisted of treadmill running sessions (1 h/day, 5 days/wk, 50–60%VO2max). The PE adipocyte size analysis revealed significant differences between the groups. The adipocyte cross-sectional area (in µm2) was significantly larger in D than in the T and S groups (3,474 ± 68.8; 1,945.7 ± 45.6; 2,492.4 ± 49.08, respectively, P < 0.05). Compared with T, the isolated adipose cells (of the D rats) showed a 48% increase in the ability to perform lipogenesis (both basal and maximally insulin-stimulated) and isoproterenol-stimulated lipolysis. No changes were observed with respect to unstimulated lipolysis. A 15% reduction in the proportion of apoptotic adipocytes was observed in groups T and D compared with group S. The gene expression levels of adiponectin and PPAR-gamma were upregulated by factors of 3 and 2 in D vs. S, respectively. PREF-1 gene expression was 3-fold higher in T vs. S. From these results, we hypothesize that adipogenesis was stimulated in group D and accompanied by significant adipocyte hypertrophy and an increase in the lipogenic capacity of the adipocytes. The occurrence of apoptotic nuclei in PE fat cells was reduced in the D and T rats; these results raise the possibility that the adipose tissue changes after detraining are obesogenic.