Plasma kinetics of an LDL-like nanoemulsion and lipid transfer to HDL in subjects with glucose intolerance

OBJECTIVE: Glucose intolerance is frequently associated with an altered plasma lipid profile and increased cardiovascular disease risk. Nonetheless, lipid metabolism is scarcely studied in normolipidemic glucose-intolerant patients. The aim of this study was to investigate whether important lipid metabolic parameters, such as the kinetics of LDL free and esterified cholesterol and the transfer of lipids to HDL, are altered in glucose-intolerant patients with normal plasma lipids. METHODS: Fourteen glucose-intolerant patients and 15 control patients were studied; none of the patients had cardiovascular disease manifestations, and they were paired for age, sex, race and co-morbidities. A nanoemulsion resembling a LDL lipid composition (LDE) labeled with 14C-cholesteryl ester and ³H-free cholesterol was intravenously injected, and blood samples were collected over a 24-h period to determine the fractional clearance rate of the labels by compartmental analysis. The transfer of free and esterified cholesterol, triglycerides and phospholipids from the LDE to HDL was measured by the incubation of the LDE with plasma and radioactivity counting of the supernatant after chemical precipitation of non-HDL fractions. RESULTS: The levels of LDL, non-HDL and HDL cholesterol, triglycerides, apo A1 and apo B were equal in both groups. The 14C-esterified cholesterol fractional clearance rate was not different between glucose-intolerant and control patients, but the ³H-free-cholesterol fractional clearance rate was greater in glucose-intolerant patients than in control patients. The lipid transfer to HDL was equal in both groups. CONCLUSION: In these glucose-intolerant patients with normal plasma lipids, a faster removal of LDE free cholesterol was the only lipid metabolic alteration detected in our study. This finding suggests that the dissociation of free cholesterol from lipoprotein particles occurs in normolipidemic glucose intolerance and may participate in atherogenic signaling

Plasma kinetics of an LDL-like nanoemulsion and lipid transfer to HDL in subjects with glucose intolerance

OBJECTIVE: Glucose intolerance is frequently associated with an altered plasma lipid profile and increased cardiovascular disease risk. Nonetheless, lipid metabolism is scarcely studied in normolipidemic glucose-intolerant patients. The aim of this study was to investigate whether important lipid metabolic parameters, such as the kinetics of LDL free and esterified cholesterol and the transfer of lipids to HDL, are altered in glucose-intolerant patients with normal plasma lipids. METHODS: Fourteen glucose-intolerant patients and 15 control patients were studied; none of the patients had cardiovascular disease manifestations, and they were paired for age, sex, race and co-morbidities. A nanoemulsion resembling a LDL lipid composition (LDE) labeled with C-14-cholesteryl ester and H-3-free cholesterol was intravenously injected, and blood samples were collected over a 24-h period to determine the fractional clearance rate of the labels by compartmental analysis. The transfer of free and esterified cholesterol, triglycerides and phospholipids from the LDE to HDL was measured by the incubation of the LDE with plasma and radioactivity counting of the supernatant after chemical precipitation of non-HDL fractions. RESULTS: The levels of LDL, non-HDL and HDL cholesterol, triglycerides, apo A1 and apo B were equal in both groups. The 14 C-esterified cholesterol fractional clearance rate was not different between glucose-intolerant and control patients, but the H-3-free- cholesterol fractional clearance rate was greater in glucose-intolerant patients than in control patients. The lipid transfer to HDL was equal in both groups. CONCLUSION: In these glucose-intolerant patients with normal plasma lipids, a faster removal of LDE free cholesterol was the only lipid metabolic alteration detected in our study. This finding suggests that the dissociation of free cholesterol from lipoprotein particles occurs in normolipidemic glucose intolerance and may participate in atherogenic signaling.This study was supported by Fundação do Amparo à Pesquisa do Estado\ud de São Paulo (FAPESP), São Paulo, Brazil. Dr. Maranhão has a Research\ud Award from Conselho Nacional de Desenvolvimento Científico e\ud Tecnológico (CNPq) Brasília, Brazil. Dr. Bertato had a scholarship from\ud CNPq

Karyological study of Ololygon tripui (Lourenço, Nascimento and Pires, 2009), (Anura, Hylidae) with comments on chromosomal traits among populations

To increase the number of cytogenetic characters used in Ololygon tripui systematics, we applied some cytogenetic techniques such as Giemsa, C- and NOR-banding, and fluorescence in situ hybridization (FISH) with 18S rDNA and repetitive microsatellite DNA probes to the study of four populations from Minas Gerais State (southeastern Brazil). All populations showed 2n = 24 and FN = 48, and chromosomal formula 8m + 10sm + 6st. Nucleolar organizing regions (NORs) were located on chromosome pair 6 in all populations, although in the Tripuí locality additional markings were observed on one homologue of chromosome pair 3. These patterns were partially congruent with results obtained using the 18S rDNA FISH probe. The microsatellites repetitive DNA (GA) 15 and (CAT) 10 probes accumulated predominantly in the terminal region of all chromosomes. Chromosome morphology and Ag-NOR were conserved among populations, a conserved pattern in Ololygon Fitzinger, 1843. Repetitive DNA FISH probes patterns were similar among populations, but they revealed species-specific differences when compared with other species of the genus Ololygon, suggesting that molecular cytogenetics are potentially more informative in karyologically conservative taxa

Two-dimensional PCA highlights the differentiated antitumor and antimicrobial activity of methanolic and aqueous extracts of Laurus nobilis L. from different origins

"Article ID 520464"Natural matrices are important sources of new antitumor and antimicrobial compounds. Species such as Laurus nobilis L. (laurel) might be used for this purpose, considering its medicinal properties. Herein, in vitro activity against human tumor cell lines, bacteria, and fungi was evaluated in enriched phenolic extracts. Specifically, methanol and aqueous extracts of wild and cultivated samples of L. nobilis were compared considering different phenolic groups. Principal component analysis (PCA) was applied to understand how each extract acts differentially against specific bacteria, fungi, and selected human tumor cell lines. In general, the extract type induced the highest differences in bioactivity of laurel samples. However, from the PCA biplot, it became clear that wild laurel samples were higher inhibitors of tumor cell lines (HeLa, MCF7, NCI-H460, and HCT15). HepG2 had the same response to laurel from wild and cultivated origin. It was also observed that methanolic extracts tended to have higher antimicrobial activity, except against A. niger, A. fumigatus, and P. verrucosum. The differences in bioactivity might be related to the higher phenolic contents in methanolic extracts. These results allow selecting the extract type and/or origin with highest antibacterial, antifungal, and antitumor activity.The authors are grateful to Fundacao para a Ciencia e a Tecnologia (FCT, Portugal) for the financial support to CIMO (strategic project PEst-OE/AGR/UI0690/2011) and REQUIMTE (PEst-C/EQB/LA0006/2011). M. I. Dias, R. Calhelha, and J. C. M. Barreira also thank FCT, POPHQREN, and FSE for their Grants (SFRH/BD/84485/2012, SFRH/BPD/ 68344/2010, and BPD/72802/2010). The authors also thank to Serbian Ministry of Education and Science for financial support (grant number 173032)

O projecto nacional de educação pelos pares da Fundação Portuguesa “A comunidade contra a sida” em escolas EB2/3 do Porto

O Projecto Nacional de Educação pelos Pares, focado na sexualidade e prevenção do VIH/ SIDA, foi criado pela Fundação Portuguesa “A Comunidade Contra a SIDA” no âmbito do Centro de Aconselhamento e Orientação de Jovens (CAOJ). O processo de aprendizagem, desenvolvido a partir do diagnóstico das necessidades de formação do público-alvo, recorre a metodologias activas que visam educar os adolescentes para uma vivência gratificante da sua sexualidade e prepará-los para assumirem o seu papel de “pares educadores” de colegas mais jovens. Esta educação de pares dirigida a alunos que frequentam o 3º ciclo do Ensino Básico é dinamizada, numa primeira fase, por jovens voluntários universitários organizados em Brigadas Universitárias de Intervenção (BUI) e no Teatro Universitário de Intervenção (TUI). Estes voluntários, com o acompanhamento de professores destacados para o CAOJ, responsáveis pela sua formação pedagógica, desenvolvem o projecto com alunos do 7º ao 9º ano de escolaridade. No terceiro ano de formação, os alunos do 3º ciclo organizados em Brigadas Escolares de Intervenção (BEI) iniciam a sua acção como educadores dos colegas mais novos, geralmente dos 2º e 1º ciclos. Esta comunicação, pretende mostrar a dinâmica deste Projecto desenvolvido em cinco escolas EB2/3 do Porto e discutir, com base em evidências recolhidas junto dos professores, voluntários e alunos envolvidos, alguns resultados obtidos neste ano lectivo

Projecto de educação pelos pares em escolas do Porto durante o ano lectivo 2009/2010

O Projecto Nacional de Educação pelos Pares, criado pela Fundação Portuguesa “A Comunidade Contra a Sida” visa a prevenção do VIH/SIDA e outros comportamentos de risco, através do desenvolvimento de projectos educativos implementados por voluntários universitários nos 7º e 8º anos de escolaridade. Estes estudantes universitários foram formados científica e pedagogicamente pela Fundação, e implementaram o projecto nacional de educação pelos pares com a supervisão de professores destacados para a FPCCS. Quando os alunos que iniciaram este projecto no 7º ano de escolaridade (12-13 anos) chegam ao 9º ano (14-15 anos), educam os seus pares dos 1º e 2º ciclos (6-11 anos) do ensino básico. A presente comunicação pretende discutir alguns dados recolhidos em seis escolas do porto envolvidas no Projecto Nacional de Educação pelos Pares, através de dois questionários de auto-resposta, designados "Sexualidade e SIDA", implementados no início do 1º e 2º anos de intervenção, que visam avaliar as necessidades de formação dos alunos nesta área. Os resultados obtidos mostraram que alguns alunos das escolas ignoram como prevenir a infecção pelo VIH e quais são os seus meios de transmissão. Por outro lado, têm a noção de que a infecção pelo VIH está associada a 'comportamentos de risco' não a 'grupos de risco'. A análise dos questionários do 8º ano evidenciou que os alunos gostaram mais de realizar dinâmicas de grupo, pelo impacto que tiveram na sua assertividade e na capacidade para reflectir sobre as consequências das suas escolhas e das decisões que tomam em relação ao seu comportamento sexual

Removal from the plasma of the free and esterified forms of cholesterol and transfer of lipids to HDL in type 2 diabetes mellitus patients

Background: The aim was to investigate new markers for type 2 diabetes (T2DM) dyslipidemia related with LDL and HDL metabolism. Removal from plasma of free and esterified cholesterol transported in LDL and the transfer of lipids to HDL are important aspects of the lipoprotein intravascular metabolism. The plasma kinetics (fractional clearance rate, FCR) and transfers of lipids to HDL were explored in T2DM patients and controls, using as tool a nanoemulsion that mimics LDL lipid structure (LDE). Results: C-14- cholesteryl ester FCR of the nanoemulsion was greater in T2DM than in controls (0.07 +/- 0.02 vs. 0.05 +/- 0.01 h(-1), p = 0.02) indicating that LDE was removed faster, but FCR H-3- cholesterol was equal in both groups. Esterification rates of LDE free-cholesterol were equal. Cholesteryl ester and triglyceride transfer from LDE to HDL was greater in T2DM (4.2 +/- 0.8 vs. 3.5 +/- 0.7%, p = 0.03 and 6.8 +/- 1.6% vs. 5.0 +/- 1.1, p = 0.03, respectively). Phospholipid and free cholesterol transfers were not different. Conclusions: The kinetics of free and esterified cholesterol tended to be independent in T2DM patients and the lipid transfers to HDL were also disturbed. These novel findings may be related with pathophysiological mechanisms of diabetic macrovascular disease.This study was supported by Fundação do Amparo à Pesquisa do Estado de\ud São Paulo (FAPESP), São Paulo, Brazil. Dr. Maranhão has a Research Award\ud from Conselho Nacional de Desenvolvimento Científico e Tecnológico\ud (CNPq) Brasília, Brazil

Relationship between latent and rebound viruses in a clinical trial of anti-HIV-1 antibody 3BNC117.

A clinical trial was performed to evaluate 3BNC117, a potent anti-HIV-1 antibody, in infected individuals during suppressive antiretroviral therapy and subsequent analytical treatment interruption (ATI). The circulating reservoir was evaluated by quantitative and qualitative viral outgrowth assay (Q2VOA) at entry and after 6 mo. There were no significant quantitative changes in the size of the reservoir before ATI, and the composition of circulating reservoir clones varied in a manner that did not correlate with 3BNC117 sensitivity. 3BNC117 binding site amino acid variants found in rebound viruses preexisted in the latent reservoir. However, only 3 of 217 rebound viruses were identical to 868 latent viruses isolated by Q2VOA and near full-length sequencing. Instead, 63% of the rebound viruses appeared to be recombinants, even in individuals with 3BNC117-resistant reservoir viruses. In conclusion, viruses emerging during ATI in individuals treated with 3BNC117 are not the dominant species found in the circulating latent reservoir, but frequently appear to represent recombinants of latent viruses

Insights into milk-clotting activity of latex peptidases from <i>Calotropis procera</i> and <i>Cryptostegia grandiflora</i>

Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl2. However, pre-incubation of both samples at 75 °C for 10 min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16 kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.Centro de Investigación de Proteínas Vegetale

Insights into milk-clotting activity of latex peptidases from <i>Calotropis procera</i> and <i>Cryptostegia grandiflora</i>

Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl2. However, pre-incubation of both samples at 75 °C for 10 min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16 kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.Centro de Investigación de Proteínas Vegetale