In hemodialysis, adiponectin, and pro-brain natriuretic peptide levels may be subjected to variations in body mass index

Adiponectin exerts cardiovascular protective actions, although some studies have shown the opposite. In hemodialysis, obese subjects display lower mortality rates despite hypoadiponectinemia, while higher adiponectin concentrations correlate with an elevated cardiovascular risk in nonobese subjects. The aim of the study is to suggest that adiponectin level variations are associated with differences in the body mass index (BMI). The interplay between adiponectin and pro-brain natriuretic peptide (Pro-BNP) levels may vary according to body fat mass. Fifty-two chronic hemodialysis patients were divided into three groups. Group A, BMI<25 (n=20); Group B, BMI 25 to 30 (n=21), and Group C, BMI>30 (n=11). Diabetics: Group A 10%; Group B 6 29%; Group C 55%, P=0.027. Determinations: Adiponectin, Pro-BNP, insulin, insulin resistance (HOMA), troponin T, nutritional status, ultrafiltration rates, C-reactive protein (CRP), vascular accesses, and echocardiography. Group A: adiponectinemia positively and significantly correlated with Pro-BNP, CRP, and troponin T. As BMI increased, adiponectin, Pro-BNP, and malnutrition significantly decreased, while insulin, HOMA, and ultrafiltration rates significantly increased. Cardiac restriction was significantly higher in obese patients. In all groups, Pro-BNP and troponin T displayed a strong positive correlation. In low-BMI subjects, high Pro-BNP and adiponectin, low myocardial restriction, and worse nutritional status were prevalent. In obesity, hypoadiponectinemia stimulates cardiac remodeling, cardiac hypertrophy, and decreased stretching, rendering Pro-BNP levels low despite high ultrafiltration rates. Thus, adiponectin correlates inversely with BMI, probably playing different cardiovascular roles as BMI changes.     

Candida albicans Tpk1p and Tpk2p isoforms differentially regulate pseudohyphal development, biofilm structure, cell aggregation and adhesins expression

Candida albicans undergoes a reversible morphological transition from single yeast cells to pseudohyphal and hyphal filaments. In this organism, cAMP-dependent protein kinase (PKA), coded by two catalytic subunits (TPK1 and TPK2) and one regulatory subunit (BCY1), mediates basic cellular processes, such as the yeast-to-hypha transition and cell cycle regulation. It is known that both Tpk isoforms play positive roles in vegetative growth and filamentation, although distinct roles have been found in virulence, stress response and glycogen storage. However, little is known regarding the participation of Tpk1p and/or Tpk2p in pseudohyphal development. This point was addressed using several C. albicans PKA mutants having heterozygous or homozygous deletions of TPK1 and/or TPK2 in different BCY1 genetic backgrounds. We observed that under hypha-only inducing conditions, all BCY1 heterozygous strains shifted growth toward pseudohyphal morphology; however, the pseudohypha:hypha ratio was higher in strains devoid of TPK2. Under pseudohypha-only inducing conditions, strains lacking TPK2 were prone to develop short and branched pseudohyphae. In tpk2 Δ/tpk2 Δ strains, biofilm architecture was markedly less dense, composed of short pseudohyphae and blastospores with reduced adhesion ability to abiotic material, suggesting a significant defect in cell adherence. Immunolabelling assays showed a decreased expression of adhesins Als1p and Als3p only in the tpk2 Δ/tpk2 Δ strain. Complementation of this mutant with a wild-type copy of TPK2 restored all the altered functions: pseudohyphae elongation, biofilm composition, cell aggregation and adhesins expression. Our study suggests that the Tpk2p isoform may be part of a mechanism underlying not only polarized pseudohyphal morphogenesis but also cell adherence.     

Diamondoid Amino Acid-Based Peptide Kinase A Inhibitor Analogues

The incorporation of diamondoid amino acids (DAAs) into peptide-like drugs is a general strategy to improve lipophilicity, membrane permeability, and metabolic stability of peptidomimetic pharmaceuticals. We designed and synthesized five novel peptidic DAA-containing kinase inhibitors of protein kinase A using a sophisticated molecular dynamics protocol and solid-phase peptide synthesis. By means of a thermophoresis binding assay, NMR, and crystal structure analysis, we determined the influence of the DAAs on the secondary structure and binding affinity in comparison to the native protein kinase inhibitor, which is purely composed of proteinogenic amino acids. Affinity and binding pose are largely conserved. One variant showed 6.5-fold potency improvement, most likely related to its increased side chain lipophilicity. A second variant exhibited slightly decreased affinity presumably due to loss of hydrogen-bond contacts to surrounding water molecules of the first solvation shell.     

Azorean macroalgae (Petalonia binghamiae,Halopteris scoparia and Osmundea pinnatifida) bioprospection: a study of fatty acid profiles and bioactivity

The Azorean macroalgae Petalonia binghamiae, Halopteris scoparia and Osmundea pinnatifida are undervalued and require further study regarding their potential use as food. These three seaweed species had low fat contents, and their fatty acid (FA) profiles were characterised by a high proportion of saturated FA (SFA) in the case of O. pinnatifida and similar weights of SFA and polyunsaturated FA (PUFA) in the other two species. Within the PUFAs, the ω3 PUFAs were the most prominent in O. pinnatifida and P. binghamiae, yielding ω3/ω6 ratios that were higher than one. Eicosapentaenoic acid (EPA, 20:5 ω3) was the most important ω3 PUFA (10–14%). Regarding the other bioactive compounds, the polyphenols were found to reach non-negligible levels (140–220 mg/100 g dw). Significant antioxidant activity was detected. Ethanolic extracts of H. scoparia and aqueous extracts of P. binghamiae showed cyclooxygenase-2 (COX-2) inhibitory capacities between 40% and 79%, indicating anti-inflammatory activity.     

Phenolic profiles of andean mashua (Tropaeolum tuberosum Ruíz & Pavón) tubers: Identification by HPLC-DAD and evaluation of their antioxidant activity

Qualitative high performance liquid chromatography with diode array detection (HPLC-DAD) was performed to characterize non-anthocyanin phenolic compounds in three different coloured mashua genotypes. The ORAC antioxidant activity contribution in the tubers related to the type of phenolic compounds present was also evaluated. Phenolic compounds were analysed by separating them into four main fractions: fraction I obtained by means of a liquid–liquid partition with ethyl acetate and fractions II, III and IV obtained by elution on a Sephadex LH-20 column. Fraction I revealed the presence of gallic acid, gallocatechin, procyanidin B₂ and epigallocatechin. Other phenolic compounds such as hydroxycinnamic and hydroxybenzoic acid derivatives, rutin and/or myricetin derivatives were also present in fraction I. Fraction II was mainly composed of epicatechin, hydroxycinnamic and hydroxybenzoic acid derivatives. Fraction III presented mainly anthocyanins for the purple coloured mashua tubers and rutin, hydroxycinnamic acid and hydroxybenzoic acid derivatives for the yellow coloured genotype. Fraction IV was composed of proanthocyanidins. Alkaline and acid hydrolysis of the different fractions revealed the presence of gallocatechin, epicatechin, p-coumaric acid, o-coumaric acid, cinnamic acid, protocatechuic acid, rutin and quercetin as the main phenolic moieties present. The proanthocyanidin fractions were the major contributors to the ORAC antioxidant activity of the mashua tubers for two of the three genotypes (34.7–39.2%). The results obtained in the present study confirm that mashua tubers constitute a promising source of antioxidant phenolics and could potentially be considered as a functional food with beneficial health effects.     

Antioxidant properties of mashua (Tropaeolum tuberosum) phenolic extracts against oxidative damage using biological in vitro assays

Purified mashua extracts (PME) from four different coloured mashua genotypes were assayed for oxidative damage prevention. Three in vitro assays for oxidative damage to biological structures rich in polyunsaturated fatty acids (PUFA), such as LDL and erythrocytes, were tested: AAPH-induced TBARS assay and Cu²⁺-induced conjugated dienes assay for LDL oxidation and AAPH-induced oxidative hemolysis of erythrocytes. Additionally, ORAC antioxidant capacity, total phenolics (TP), total flavanoids (TFA) and total anthocyanins (TA) were evaluated. In the presence of 5 μM of gallic acid equivalents (GAE), inhibitions of LDL oxidation for the PME ranged from 29.1% to 34.8% and from 51.8% to 58.1% when the TBARS and conjugated dienes assays were performed, respectively. PME inhibited the hemolysis of erythrocytes within the range 20.8–25.1%. Thus, mashua phenolic extracts are capable of scavenging peroxyl radicals, as well as chelating redox metal ions in vitro. ORAC and LDL protection (TBARS and conjugated dienes assays) showed good correlations with the TP and TFA, suggesting that these compounds have a good ability to protect LDL molecules under the employed conditions. In contrast, inhibition of hemolysis did not show any correlation with the evaluated phenolic assays (TP, TA, TFA) or with any of the evaluated oxidative LDL assays, suggesting a specific action of some non-evaluated compounds present in the PME. The results of this study indicate that the mashua polyphenol extracts displayed good antioxidant properties against oxidative damage in biological structures rich in PUFA. The displayed antioxidant properties could be applied in the field of food or cosmetic industry.     

Structural investigation of the HIV-1 envelope glycoprotein gp160 cleavage site, 2: relevance of an N-terminal helix

Proteolytic activation of the HIV-1 envelope glycoprotein gp160 is selectively performed by the proprotein convertase furin at the C-terminus of the sequence R508-E-K-R511 (site 1), in spite of the presence of another consensus sequence, Lys500-Ala-Lys-Arg503 (site 2). On the basis of the solution structural analysis of the synthetic peptide p498, spanning the gp160 sequence Pro498-Gly516, we previously suggested a possible role of an N-terminal helix in regulating the exposure and accessibility of the gp160 physiological cleavage site, enclosed in a loop. Here we report on the activity and conformation of the 23-residue peptide h-REKR, designed to exhibit a large N-terminal helix, followed by the gp160 native sequence, Arg508-Gly516. h-REKR is digested by furin with high efficiency, comparable to the full native p498. Circular dichroism analyses, in mixtures from pure water to 98 % trifluoroethanol, outline a significant content of helical structure in the peptide conformation. The molecular model obtained from NMR data collected in trifluoroethanol/water, by means of DYANA and AMBER simulations, indeed has helical structure on a large N-terminal segment. Such a long helix does not seem to affect the loop conformation of the C-terminal site 1-containing sequence, which exhibits the same proton chemical shifts already observed for the full native p498.     

An algorithm for predicting proton nuclear magnetic resonance deshielding over a carbon-carbon double bond

Hydrogen nuclei located over a carbon-carbon double bond in a strong magnetic field experience NMR shielding effects that result from the magnetic anisotropy of the nearby double bond and various other intramolecular shielding effects. We have used GIAO, a subroutine in Gaussian 98, to calculate isotropic shielding values and to predict the proton NMR shielding increment for a simple model system: methane held in various orientations, positions, and distances over ethene. The average proton NMR shielding increments of several orientations of methane have been plotted versus the Cartesian coordinates of the methane protons relative to the center of ethene. A single empirical equation for predicting the NMR shielding experienced by protons over a carbon-carbon double bond has been developed from these data. The predictive capability of this equation has been validated by comparing the shielding increments for several alkenes calculated using our equation to the experimentally observed shielding increments. This equation predicts the NMR shielding effects more accurately than previous models that were based on fewer geometries of methane over ethene. In fact, deshielding is predicted by this equation for protons over the center and within about 3 A of a carbon-carbon double bond. This result is in sharp contrast to predictions made by the long-held McConnell "shielding cone" model found in nearly every textbook on NMR, but is consistent with experimental observations. The algorithm for predicting the (de)shielding increment for a proton over an alkene can be used in a spreadsheet on a PC or incorporated into software that estimates chemical shifts using additive substituent constants or a database of structures. In either application its use can substantially improve the accuracy of the estimated chemical shift of a proton in the vicinity of a carbon-carbon double bond, and thus assist in spectral assignments and in correct structure determination.     

Acceleration of osmotic dehydration process through ohmic heating of foods: raspberries (Rubus idaeus)

Raspberries (Rubus idaeus) were osmotically dehydrated by applying a conventional method under the supposition of a homogeneous solution, all in a 62% glucose solution at 50 degrees C. Raspberries (Rubus idaeus) were also osmotically dehydrated by using ohmic heating in a 57% glucose solution at a variable voltage (to maintain temperature between 40 and 50 degrees C) and an electric field intensity <100 V/cm. When comparing the results from both experiments it was evident that processing time is reduced when ohmic heating technique was used. In some cases this reduction reached even 50%. This is explained by the additional effect to the thermal damage that is generated in an ohmic process, denominated electroporation.