AKT and AMPK Activation after High-Fat and High-Glucose In Vitro Treatment of Prostate Epithelial Cells

Considering the increasing consumption of saturated fat and glucose in diets worldwide and its possible association to carcinogenesis, this investigation analysed the proliferation profile of nonmalignant human prostate epithelial cells after exposure to elevated levels of fat and glucose. PNT1A cells were cultured with palmitate (100 or 200 mu M) and/or glucose (450mg/dl) for 24 or 48 h. Treated cells were evaluated for viability test and cell proliferation (MTS assay). AKT and AMPK phosphorylation status were analysed by Western blotting. After 24 h of high-fat alone or associated with high-glucose treatment, there was an increase in AMPK and AKT activation associated to unchanged MTS-cell proliferation. Following 48 h of high-fat but not high-glucose alone, cells decreased AMPK activation and maintained elevated AKT levels. These data were associated to increased cell proliferation after further high-fat treatment. After longer high-fat exposure, MTS revealed that cells remained proliferating. High-glucose alone or associated to high-fat treatment was not able to increase cell proliferation and AKT activation. A high-fat medium containing 100 mu M of palmitate stimulates proliferation in PNT1A cells by decreasing the activation of AMPK and increasing activation of AKT after longer exposure time. These findings improve the knowledge about the negative effect of high levels of this saturated fatty acid on proliferative disorders of prostate.

Pancreatic Alpha-Cell Dysfunction Contributes to the Disruption of Glucose Homeostasis and Compensatory Insulin Hypersecretion in Glucocorticoid-Treated Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Apical periodontitis and periodontal disease increase serum IL-17 levels in normoglycemic and diabetic rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preventive Effects of Chitosan Coacervate Whey Protein on Body Composition and Immunometabolic Aspect in Obese Mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hematological and biochemical changes in a Phrynops geoffroanus (Schweigger, 1812) population in response to environmental stressors

Turtles are among the most endangered vertebrate groups, and the main threats to populations are environmental pollution and habitat degradation. The species Phrynops geoffroanus, popularly known as “Geoffroy’s side-necked turtle”, has proliferated in polluted environments, where adverse conditions could influence their living habits and physiological condition. Studies that monitor the effects of environmental pollution are key to understanding the species’ biology and designing effective conservation strategies. Thus, the analysis of hematological and biochemical parameters has been shown to be important in assessing the health of wild animals and risks for the animal and ecosystem. This study aimed to assess the environmental influence on the physiology of a P. geoffroanus population through the evaluation of antioxidant status and responses to environmental stressors, compared to specimens from a place under controlled conditions. Blood samples of 60 specimens were collected, 30 from the Felicidade Stream, polluted environment, within the city of São José do Rio Preto, and 30 from the “Reginaldo Uvo Leone” breeding farm, Tabapuã, SP, a place under controlled conditions, whose samples constituted the control group. They were evaluated by hemogram and by determining thiobarbituric acid reactive species (TBARS), Trolox-equivalent antioxidant capacity (TEAC) and the activities of the antioxidant enzymes catalase and glucose-6-phosphate dehydrogenase (G6PDH). There was a wide variation in hematological parameters of P. geoffroanus from the urban environment. The red blood cell count and hemoglobin values were significantly less than those observed in animals from the breeding farm (P = 0.0004; P = 0.0371, respectively). There was a significant increase in the number of thrombocytes (P < 0.0001) and leukocytes (P < 0.0001) in the animals from Felicidade Stream. The stress indices were similar between the two groups (P = 0.4077). TBARS levels showed the cytotoxic potential of compounds in the urban environment, whose animals had elevated levels of lipid peroxidation (P < 0.0001), despite showing a response to environmental damages with increase in antioxidant capacity, as demonstrated by the TEAC assay (P = 0.0207). The lower catalase enzyme activity noted in individuals from the urban environment (P = 0.000184) could be due to the presence of inhibitory compounds. On the other hand, G6PDH activity was higher (P = 0.002962), where this enzyme acts in the generation of NADPH, which is used in several detoxification pathways. We conclude that environmental contamination can increase oxidative damages and generate physiological changes in this species. These data are very useful for the conservation of P. geoffroanus and turtles in general, and confirm that these techniques are effective in monitoring natural regions and that P. geoffroanus can serve as an environmental contamination bioindicator.

Social challenge increases cortisol and hypothalamic monoamine levels in matrinxã (Brycon amazonicus)

The neural circuitry for social behavior and aggression appears to be evolutionarily conserved across the vertebrate subphylum and involves a complex neural network that includes the hypothalamus as a key structure. In the present study, we evaluated the changes in monoamine levels in the hypothalamus and on serum cortisol and plasma glucose of resident matrinxã (Brycon amazonicus) submitted to a social challenge (introduction of an intruder in their territory). The fight promoted a significant increase in hypothalamic 5-HT, NA and DA levels and on the metabolites 5-HIAA and DOPAC, and decreased 5-HIAA/5-HT and DOPAC/DA ratios in resident fish. Furthermore, an increase in serum cortisol and plasma glucose was also observed after the fight. Resident fish presented a high aggressiveness even with increased 5-HT levels in the hypothalamus. The alteration in hypothalamic monoaminergic activity of matrinxã suggests that this diencephalic region is involved in aggression and stress modulation in fish; however, it does not exclude the participation of other brain areas not tested here.

Investigation of cytokines, oxidative stress, metabolic, and inflammatory biomarkers after orange juice consumption by normal and overweight subjects

Abdominal adiposity has been linked to metabolic abnormalities, including dyslipidemia, oxidative stress, and low-grade inflammation. To test the hypothesis that consumption of 100% orange juice (OJ) would improve metabolic, oxidative, and inflammatory biomarkers and cytokine levels in normal and overweight subjects with increased waist circumference. Subjects were divided into two groups in accordance with their body mass index: normal and overweight. Both groups of individuals consumed 750 mL of OJ daily for 8 weeks. Body composition (weight, height, percentage of fat mass, and waist circumference); metabolic biomarkers (total cholesterol, low-density lipoprotein-cholesterol [LDL-C], high-density lipoprotein-cholesterol [HDL-C], triglycerides, glucose, insulin, HOMA-IR, and glycated hemoglobin); oxidative biomarkers (malondialdehyde and DPPH(•)); inflammatory biomarkers (high-sensitivity C-reactive protein [hsCRP]); cytokines (IL-4, IL-10, IL-12, TNF-α, and IFN-γ); and diet were evaluated before and after consumption of OJ for 8 weeks. The major findings of this study were: 1) no alteration in body composition in either group; 2) improvement of the lipid profile, evidenced by a reduction in total cholesterol and LDL-C; 3) a potential stimulation of the immune response due to increase in IL-12; 4) anti-inflammatory effect as a result of a marked reduction in hsCRP; and 5) antioxidant action by the enhancement of total antioxidant capacity and the reduction of lipid peroxidation, in both normal and overweight subjects. OJ consumption has a positive effect on important biomarkers of health status in normal and overweight subjects, thereby supporting evidence that OJ acts as functional food and could be consumed as part of a healthy diet to prevent metabolic and chronic diseases.

Alcohol levels in cerebrospinal fluid and blood samples from patients under pathological conditions

We measured alcohol levels by the Cordebard method in 148 CSF samples from individuals who had abstained from alcohol for at least 7 days prior to the beginning of the study. Each blood sample was accompanied by a CSF sample from the same patient. CSF samples found to be normal after analysis were used as controls. Mean alcohol concentration in blood did not differ significantly between the control group and the groups with altered CSF. The group with altered CSF had statistically higher alcohol levels in CSF than in blood. CSF lactate, glucose and protein levels were not correlated with alcohol level. The results suggest the presence of endogenous alcohol in the CSF, with levels increasing in the presence of pathological processes involving the nervous system.

Daily intake of pasteurized orange juice decreases serum cholesterol, fasting glucose and diastolic blood pressure in adults

Previous studies have shown that fresh squeezed orange juice or juice from reconstituted concentrate can help prevent the development of atherosclerosis. Pasteurized orange juice presently represents the major orange juice available in the market, and because of this, it becomes necessary to determine the healthy benefits associated with this product. In this study we investigated the effect of regular consumption of pasteurized orange juice on the nutritional status, biochemical profile, and arterial blood pressure in healthy men and women. Men and women volunteered to consume pasteurized orange juice (500 mL·d–1 and 750 mL·d–1, respectively), for 8 weeks. Anthropometric, biochemical, hemodynamic, and dietary assessments were evaluated at baseline and at the end of the experimental period. Total cholesterol and LDL-C significantly decreased in both men and women after the consumption of orange juice, and an increase in HDL-C level was detected exclusively in women. Fasting glucose, diastolic blood pressure, and triglyceride levels dropped in men after the consumption of orange juice. Anthropometric variables did not change with orange juice consumption, only waist circumference decreased significantly in women. Consumption of orange juice increased the energy and carbohydrate intake for women; however, vitamin C and folate increased after the orange juice period for both men and women. Regular consumption of pasteurized orange juice by men (750 mL·d–1) and women (500 mL·d–1) reduced the risk of developing atherosclerosis, and increased the nutritional quality of their diets.

Purification and characterization of the biological effects of phospholipase A(2) from sea anemone Bunodosoma caissarum

Sea anemones contain a variety of biologically active substances. Bunodosoma caissarum is a sea anemone from the Cnidaria phylum, found only in Brazilian coastal waters. The aim of the present work was to study the biological effects of PLA(2) isolated from the sea anemone B. caissarum on the isolated perfused kidney, the arteriolar mesenteric bed and on insulin secretion. Specimens of B. caissarum were collected from the Sao Vicente Channel on the southern coast of the State of São Paulo, Brazil. Reverse phase HPLC analysis of the crude extract of B. caissarum detected three PLA(2) proteins (named BcPLA(2)1, BCPLA(2)2 and BcPLA(2)3) found to be active in B. caissarum extracts. MALDI-TOF mass spectrometry of BcPLA(2)1 showed one main peak at 14.7 kDa. The N-terminal amino acid sequence of BcPLA(2)1 showed high amino acid sequence identity with PLA(2) group III protein isolated from the Mexican lizard (PA23 HELSU, HELSU, PA22 HELSU) and with the honey bee Apis mellifera (PLA(2) and 1POC_A). In addition, BcPLA(2)1 also showed significant overall homology to bee PLA(2). The enzymatic activity induced by native BCPLA(2)1 (20 mu g/well) was reduced by chemical treatment with p-bromophenacyl bromide (p-BPB) and with morin. BcPLA(2)1 strongly induced insulin secretion in presence of high glucose concentration. In isolated kidney, the PLA(2) from B. caissarum increased the perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate, and sodium, potassium and chloride levels of excretion. BcPLA(2)1, however, did not increase the perfusion pressure on the mesenteric vascular bed. In conclusion, PLA(2), a group III phospholipase isolated from the sea anemone B. caissarum, exerted effects on renal function and induced insulin secretion in conditions of high glucose concentration. (C) 2009 Elsevier Ltd. All rights reserved.

A simple approach to integrate the ecotoxicological and chemical data for the establishment of environmental risk levels

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Acute lethal and sublethal effects of neem leaf extract on the neotropical freshwater fish Prochilodus lineatus

The aim of this study was to determine the toxicity of the aqueous extract of neem leaves, a product extensively used in fish-farms as alternative for the control of fish parasites and fish fry predators, for the neotropical fish Prochilodus lineatus. The 24 It LC(50) of neem leaf extract for juveniles P lineatus was estimated as 4.8 g L(-1); the fish were then exposed for 24 h to 2.5, 5.0 and 7.5 g L(-1) or only clean water (control). Plasma glucose levels were higher in fish exposed to 2.5 g L(-1) and 5.0 g L(-1) neem extract, relative to control, indicating a typical stress response. Neem extract did not interfere with the osmoregulating capacity of the fish, as their plasma sodium, chloride, total protein and osmolarity did not change. The presence of the biopesticide interfered with the antioxidant defense system of P. lineatus, as there was a decrease in liver catalase activity at all neem concentrations and the detoxifying enzyme glutathione-S-transferase was activated in fish exposed to 5.0 g L(-1). Fish exposed to all neem extract concentrations exhibited damaged gill and kidney tissue. These results indicate that although neem extract is less toxic to P. lineatus than other synthetic insecticides used in fish-farming it does cause functional and morphological changes in this fish species. (c) 2006 Elsevier B.V. All rights reserved.

Effects of hot and cold smoking processes on organoleptic properties, yield and composition of matrinxa fillet

Foi avaliado o efeito do processo de defumação a quente (45-90ºC/5 horas) e a frio (27-45ºC/10 horas) nas propriedades organolépticas, no rendimento e na composição dos filés de matrinxã (Brycon cephalus). Não houve diferença significativa no rendimento de filés defumados e não-defumados. As perdas no processo de defumação foram significativamente maiores para defumação a quente (19,37%) em comparação à defumação a frio (17,08%). O processo de defumação reduziu a umidade (in natura = 72,91%; defumado a quente = 58,51%; e defumado a frio = 59,68%) e aumentou os teores de proteína bruta, lipídios e cinzas. Houve diferença significativa somente nos teores de proteína no defumado a quente (28,07%) e defumado a frio (27,14%). O processo a frio resultou em melhor aparência e cor de filé, enquanto o processo a quente melhorou o sabor, o teor de sal e a aparência geral. O aroma e a textura não diferiram significativamente entre os processos. O processo de defumação a quente melhora as propriedades organolépticas e os níveis de proteína do filé de matrinxã.

Experimental Trypanosoma evansi infection in donkeys: hematological, biochemical and histopathological changes

Cinco jumentos, adultos foram infectados experimentalmente com cepa brasileira de Trypanosoma evansi, isolada de um cão naturalmente infectado, com o intuito de observar as alterações hematológicas, bioquímicas e histopatológicas durante a evolução da enfermidade. O curso da infecção experimental foi de 145 dias. Análise hematológica dos jumentos infectados revelou declínio nos valores de hemoglobina, hematócrito e contagem total de eritrócitos. Notou-se anemia após sucessivos picos de parasitemia. Análise bioquímica indicou aumento dos níveis de índice ictérico, globulinas séricas e diminuição dos valores séricos de albumina e glicose. Todos os jumentos infectados apresentaram aumento do baço e de sua polpa branca, aumento de linfonodos mediastínicos e congestão pulmonar. Meningoencefalite foi o principal achado histopatológico. em algumas áreas do pedículo cerebelar foram observadas desmielinização, além de vacuolização do neurópilo. O estudo mostrou que jumentos infectados com a cepa brasileira do T. evansi desenvolveram doença crônica.

Effects of six carbohydrate sources on diet digestibility and postprandial glucose and insulin responses in cats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)