Characterization of Primary Isolates of HIV Type 1 CRF28_BF, CRF29_BF, and Unique BF Recombinants Circulating in Sao Paulo, Brazil

We report for the first time the genetic and biological characterization of 10 HIV-1 primary isolates representing CRF28_BF and CRF29_BF together with additional unique BF recombinant forms (URFs) obtained by PBMC cocultivation. Recombination is an important factor promoting the increase in the genetic diversity of HIV-1. Notably, more than 20% of HIV-1 sequences worldwide were recombinants. Several recombinant viruses were reported in Brazil, and six circulating recombinant forms (CRFs) have been identified (CRF28_BF, CRF29_BF, CRF31_BC, CRF39_BF, CRF40_BF, and CRF46_BF). CRF28_BF and CRF29_BF were found to infect almost 30% of the patients in Sao Paulo State. The near full-length genomes of these 10 primary isolates were amplified by nested PCR in three overlapping segments, purified, and sequenced. Three samples were related to CRF28_BF, three to CRF29_BF, and four were unique recombinant forms (URFs), as determined by their breakpoint profile determined with the jpHMM program. Additionally, the coreceptor usage of these isolates was investigated in vitro using GHOST assays, which revealed three dual-tropic (X4/R5) viruses, four lymphotropic (X4) viruses, and three macrophage-tropic (R5) viruses with different V3-loop motifs, which challenges the notion that GWGR-carrying viruses are macrophage-tropic only. In sum, we report a much-anticipated well-characterized panel of viruses representing CRF28_BF, CRF29_BF, and URFs from Sao Paulo State, Brazil.

BF Integrase Genes of HIV-1 Circulating in Sao Paulo, Brazil, with a Recurrent Recombination Region

Although some studies have shown diversity in HIV integrase (IN) genes, none has focused particularly on the gene evolving in epidemics in the context of recombination. The IN gene in 157 HIV-1 integrase inhibitor-naive patients from the Sao Paulo State, Brazil, were sequenced tallying 128 of subtype B (23 of which were found in non-B genomes), 17 of subtype F (8 of which were found in recombinant genomes), 11 integrases were BF recombinants, and 1 from subtype C. Crucially, we found that 4 BF recombinant viruses shared a recurrent recombination breakpoint region between positions 4900 and 4924 (relative to the HXB2) that includes 2 gRNA loops, where the RT may stutter. Since these recombinants had independent phylogenetic origin, we argue that these results suggest a possible recombination hotspot not observed so far in BF CRF in particular, or in any other HIV-1 CRF in general. Additionally, 40% of the drug-naive and 45% of the drug-treated patients had at least 1 raltegravir (RAL) or elvitegravir (EVG) resistance-associated amino acid change, but no major resistance mutations were found, in line with other studies. Importantly, V151I was the most common minor resistance mutation among B, F and BF IN genes. Most codon sites of the IN genes had higher rates of synonymous substitutions (dS) indicative of a strong negative selection. Nevertheless, several codon sites mainly in the subtype B were found under positive selection. Consequently, we observed a higher genetic diversity in the B portions of the mosaics, possibly due to the more recent introduction of subtype F on top of an ongoing subtype B epidemics and a fast spread of subtype F alleles among the B population.

Peptidomics of Three Bothrops Snake Venoms: Insights Into the Molecular Diversification of Proteomes and Peptidomes

Snake venom proteomes/peptidomes are highly complex and maintenance of their integrity within the gland lumen is crucial for the expression of toxin activities. There has been considerable progress in the field of venom proteomics, however, peptidomics does not progress as fast, because of the lack of comprehensive venom sequence databases for analysis of MS data. Therefore, in many cases venom peptides have to be sequenced manually by MS/MS analysis or Edman degradation. This is critical for rare snake species, as is the case of Bothrops cotiara (BC) and B. fonsecai (BF), which are regarded as near threatened with extinction. In this study we conducted a comprehensive analysis of the venom peptidomes of BC, BF, and B. jararaca (BJ) using a combination of solid-phase extraction and reversed-phase HPLC to fractionate the peptides, followed by nano-liquid chromatography-tandem MS (LC-MS/MS) or direct infusion electrospray ionization-(ESI)-MS/MS or MALDI-MS/MS analyses. We detected marked differences in the venom peptidomes and identified peptides ranging from 7 to 39 residues in length by de novo sequencing. Forty-four unique sequences were manually identified, out of which 30 are new peptides, including 17 bradykinin-potentiating peptides, three poly-histidine-poly-glycine peptides and interestingly, 10 L-amino acid oxidase fragments. Some of the new bradykinin-potentiating peptides display significant bradykinin potentiating activity. Automated database search revealed fragments from several toxins in the peptidomes, mainly from L-amino acid oxidase, and allowed the determination of the peptide bond specificity of proteinases and amino acid occurrences for the P4-P4' sites. We also demonstrate that the venom lyophilization/resolubilization process greatly increases the complexity of the peptidome because of the imbalance caused to the venom proteome and the consequent activity of proteinases on venom components. The use of proteinase inhibitors clearly showed different outcomes in the peptidome characterization and suggested that degradomic-peptidomic analysis of snake venoms is highly sensitive to the conditions of sampling procedures. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.019331, 1245-1262, 2012.

Effects of circuit-based exercise programs on the body composition of elderly obese women

Aim: The aim of this study was to investigate the impact of circuit-based exercise on the body composition in obese older women by focusing on physical exercise and body weight (BW) gain control in older people. Methods: Seventy older women (>60 years old) voluntarily took part in the study. Participants were randomized into six different groups according to body mass index (BMI): appropriate weight (AW) control (AWC) and trained (AWT) groups, overweight (OW) control (OWC) and trained (OWT) groups, and obesity (O) control (OC) and trained (OT) groups. The exercise program consisted of 50 minutes of exercise three times per week for 12 weeks. The exercises were alternated between upper and lower body using rest between sets for 40 seconds with intensity controlled by heart rate (70% of work). The contraction time established was 5 seconds to eccentric and concentric muscular action phase. The following anthropometric parameters were evaluated: height (m), body weight (BW, kg), body fat (BF, %), fat mass (FM, kg), lean mass (LM, kg), and BMI (kg/m(2)). Results: The values (mean +/- standard deviation [SD]) of relative changes to BW (-8.0% +/- 0.8%), BF (-21.4% +/- 2.1%), LM (3.0% +/- 0.3%), and FM (-31.2% +/- 3.0%) to the OT group were higher (P < .05) than in the AWT (BW: -2.0% +/- 1.1%; BF: -4.6% +/- 1.8%; FM: -7.0% +/- 2.8%; LM: 0.2% +/- 1.1%) and OWT (BW: -4.5% +/- 1.0%; BF: -11.0% +/- 2.2%; FM: -16.1% +/- 3.2%; LM: -0.2% +/- 1.0%) groups; additionally, no differences were found for C groups. While reduction (P < .03) in BMI according to absolute values was observed for all trained groups (AWT: 22 +/- 1 versus 21 +/- 1; OWT: 27 +/- 1 versus 25 +/- 1, OT: 34 +/- 1 versus 30 +/- 1) after training, no differences were found for C groups. Conclusion: In summary, circuit-based exercise is an effective method for promoting reduction in anthropometrics parameters in obese older women.

Electrostatic potentials and polarization effects in proton-molecule interactions by means of multipoles from the quantum theory of atoms in molecules

Some atomic multipoles (charges, dipoles and quadrupoles) from the Quantum Theory of Atoms in Molecules (QTAIM) and CHELPG charges are used to investigate interactions between a proton and a molecule (F2, Cl2, BF, AlF, BeO, MgO, LiH, H2CO, NH3, PH3, BF3, and CO2). Calculations were done at the B3LYP/6-311G(3d,3p) level. The main aspect of this work is the investigation of polarization effects over electrostatic potentials and atomic multipoles along a medium to long range of interaction distances. Large electronic charge fluxes and polarization changes are induced by a proton mainly when this positive particle approaches the least electronegative atom of diatomic heteronuclear molecules. The search for simple equations to describe polarization on electrostatic potentials from QTAIM quantities resulted in linear relations with r-4 (r is the interaction distance) for many cases. Moreover, the contribution from atomic dipoles to these potentials is usually the most affected contribution by polarization what reinforces the need for these dipoles to a minimal description of purely electrostatic interactions. Finally, CHELPG charges provide a description of polarization effects on electrostatic potentials that is in disagreement with physical arguments for certain of these molecules. (c) 2012 Wiley Periodicals, Inc.

Endothelium-Dependent Vasorelaxant Effect of Butanolic Fraction from Caryocar brasiliense Camb. Leaves in Rat Thoracic Aorta

Caryocar brasiliense Camb. "pequi" is a native plant from the Cerrado region of Brazil that contains bioactive components reported to be antioxidant agents. Previous work has demonstrated that dietary supplementation with pequi decreased the arterial pressure of volunteer athletes. We found that the crude hydroalcoholic extract (CHE) of C. brasiliense leaves relaxed, in a concentration-dependent manner, rat aortic rings precontracted with phenylephrine, and that the butanolic fraction (BF) produced an effect similar to that of the CHE. Aortic relaxation induced by BF was abolished by endothelium removal, by incubation of the nitric oxide synthase inhibitor L-NAME, or the soluble guanylatecyclase inhibitor ODQ. However, incubation with atropine and pyrilamine had no effect on the BF-induced vasorelaxation. Moreover, this effect was not inhibited by indomethacin and tetraethylammonium. The concentration-response curve to calcium in denuded-endothelium rings was not modified after incubation with BF, and the vasorelaxation by BF in endothelium-intact rings precontracted with KCl was abolished after incubation with L-NAME. In addition, administration of BF in anesthetized rats resulted in a reversible hypotension. The results reveal that C. brasiliense possesses both in vivo and in vitro activities and that the vascular effect of BF involves stimulation of the nitric oxide/cyclic GMP pathway.

Ventilatory threshold is related to walking tolerance in patients with intermittent claudication

Background: This study assessed the relationship between lower limb hemodynamics and metabolic parameters with walking tolerance in patients with intermittent claudication (IC). Patients and methods: Resting ankle-brachial index (ABI), baseline blood flow (BF), BF response to reactive hyperemia (BFRH), oxygen uptake (VO2), initial claudication distance (ICD) and total walking distance (TWD) were measured in 28 IC patients. Pearson and Spearman correlations were calculated. Results: ABI, baseline BF and BF response to RH did not correlate with ICD or TWD. VO2 at first ventilatory threshold and VO(2)peak were significantly and positively correlated with ICD (r = 0.41 and 0.54, respectively) and TWD (r = 0.65 and 0.71, respectively). Conclusions: VO(2)peak and VO2 at first ventilatory threshold, but not ABI, baseline BF and BFHR were associated with walking tolerance in IC patients. These results suggest that VO2 at first ventilatory threshold may be useful to evaluate walking tolerance and improvements in IC patients.

Stable assemblies of cationic bilayer fragments and CpG oligonucleotide with enhanced immunoadjuvant activity in vivo

The cationic lipid dioctadecyldimethylammonium bromide (DODAB) and the CpG oligonucleotide (CpG) have been separately used as potent immunoadjuvants driving Th1 responses. Here DODAB bilayer fragments (BF) and CpG (5 -TTGACGTTCG-3) assemblies have their physical properties and immunoadjuvant activity determined using ovalbumin (OVA) as a model antigen. At 0.1 mg/mL OVA, the dependence of DODAB BF/OVA size and zeta-potential on time and [DODAB] establishes 0.1 mMDODAB as suitable for obtaining stable and cationic DODAB BF/OVA assemblies. At 0.1 mMDODAB, 0.1 mg/mL OVA and 0.006 mMCpG, the zeta-potential is zero. At [CpG]>0.006 mM, good colloidal stability for the anionic assemblies is due to charge overcompensation. At 0.020 mM CpG, these DODAB BF/OVA/CpG assemblies are highly effective in vivo generating responses similar to those elicited by the stable and cationic DODAB BF/OVA. The anti-OVA DTH reaction and the secretion of IFN-gamma and IL-12 are 6, 42 and 9 times larger for the DODAB BF/OVA/CpG-immunized mice than the same responses by OVA-immunized mice, respectively. This work shows for the first time that charge of small assemblies is not important to determine the immune response. (C) 2011 Elsevier B. V. All rights reserved.

Biochemical control of prostate cancer with iodine-125 brachytherapy alone: experience from a single institution

Brachytherapy is an adequate option as monotherapy for localised prostate cancer. The objective of this study was to evaluate and compare biochemical failure free survival (BFFS) after low-dose-rate brachytherapy (LDRB) alone for patients with prostate cancer using ASTRO and Phoenix criteria, and detect prognostic factors. Data on 220 patients treated between 1998 and 2002 with LDRB were retrospectively analysed. Neoadjuvant hormone therapy was used in 74 (33.6%) patients. Median follow-up was 53.5 months (24-116). Five year BFFS was 83.0% and 83.7% using, respectively, the ASTRO and Phoenix criteria. Low -and intermediate-risk patients presented, respectively, 86.7% and 77.8% 5-year BFFS using the ASTRO definition (p=0.069), and 88.5% and 78.6% considering the Phoenix criteria (p=0.016). Bounce was observed in 66 (30%) patients. Multivariate analysis detected PSA at diagnosis < 10 ng/ml and less than 50% positive biopsy fragments as favourable prognostic factors, regarding BF using both criteria. For the Phoenix criteria, also Gleason score < 7 and low-risk group were identified as independent favourable prognostic factors. LDRB alone should be considered mostly for low-risk patients. PSA level was a strong independent prognostic factor. We support the use of the Phoenix criteria for detection of BF in patients submitted to LDRB alone.

Cassava starch biodegradable films: Influence of glycerol and clay nanoparticles content on tensile and barrier properties and glass transition temperature

In this this study, glycerol content and its incorporation method on tensile and barrier properties of biodegradable films (BF) based on cassava starch were analyzed. ANOVA showed that the glycerol incorporation method did not influence the results (P > 0.05), however the glycerol content influenced significantly the tensile and barrier properties of the films (P < 0.05). Films prepared with lower glycerol content presented better tensile and barrier properties than films with higher content. Films were then prepared with addition of clay nanoparticles and their tensile and barrier properties and glass transition temperature were measured. ANOVA indicated that both glycerol and clay nanoparticles influenced significantly the tensile and barrier properties (P < 0.05), diminishing film permeability when clay nanoparticles were present, while the glass transition temperature was not influenced (P > 0.05). (C) 2011 Elsevier Ltd. All rights reserved.

Residual stresses in Y-TZP crowns due to changes in the thermal contraction coefficient of veneers

Objective. To test the hypothesis that the difference in the coefficient of thermal contraction of the veneering porcelain above (˛liquid) and below (˛solid) its Tg plays an important role in stress development during a fast cooling protocol of Y-TZP crowns. Methods. Three-dimensional finite element models of veneered Y-TZP crowns were developed. Heat transfer analyses were conducted with two cooling protocols: slow (group A) and fast (groups B–F). Calculated temperatures as a function of time were used to determine the thermal stresses. Porcelain ˛solid was kept constant while its ˛liquid was varied, creating different ˛/˛solid conditions: 0, 1, 1.5, 2 and 3 (groups B–F, respectively). Maximum ( 1) and minimum ( 3) residual principal stress distributions in the porcelain layer were compared. Results. For the slowly cooled crown, positive 1 were observed in the porcelain, orientated perpendicular to the core–veneer interface (“radial” orientation). Simultaneously, negative 3 were observed within the porcelain, mostly in a hoop orientation (“hoop–arch”). For rapidly cooled crowns, stress patterns varied depending on ˛/˛solid ratios. For groups B and C, the patterns were similar to those found in group A for 1 (“radial”) and 3 (“hoop–arch”). For groups D–F, stress distribution changed significantly, with 1 forming a “hoop-arch” pattern while 3 developed a “radial” pattern. Significance. Hoop tensile stresses generated in the veneering layer during fast cooling protocols due to porcelain high ˛/˛solid ratio will facilitate flaw propagation from the surface toward the core, which negatively affects the potential clinical longevity of a crown.

Pressão arterial, respostas metabólicas e autonômicas à insulina e infusão de intralipid® em pacientes chagásicos

FUNDAMENTO: Infusão de intralipid e heparina resulta em aumento da pressão arterial e também em anormalidades autonômicas em indivíduos normais e hipertensos. OBJETIVO: Avaliar a sensibilidade a insulina e o impacto da infusão de intralipid e de heparina (ILH) sobre a resposta hemodinâmica, metabólica e autonômica em pacientes com a forma indeterminada da doença de Chagas. MÉTODOS: Doze pacientes com a forma indeterminada da doença de Chagas e 12 voluntários saudáveis foram avaliados. RESULTADOS: A pressão arterial basal e a frequência cardíaca foram semelhantes nos dois grupos. Os níveis plasmáticos de noradrenalina encontravam-se ligeiramente aumentados no grupo de pacientes chagásicos. Após o Teste de Tolerância a Insulina (TTI), houve um declínio significativo na glicose dos dois grupos. A Infusão de ILH resultou em aumento da pressão arterial em ambos os grupos, mas não houve nenhuma mudança significativa na noradrenalina plasmática. O componente de Baixa Frequência (BF) mostrou-se semelhante e aumentou de forma semelhante em ambos os grupos. O componente de Alta Frequência (AF) apresentou-se menor no grupo chagásico. CONCLUSÃO: Pacientes com forma indeterminada da doença de Chagas apresentaram aumento da atividade simpática no momento basal e uma resposta inadequada à insulina. Eles também tiveram um menor componente de alta frequência e sensibilidade barorreflexa prejudicada no momento basal e durante a infusão de intralipid e heparina.

Segurança da intervenção fisioterápica precoce após o infarto agudo do miocárdio

INTRODUÇÃO: A fisioterapia na fase I da reabilitação cardiovascular (FTCV) pode ser iniciada de 12 a 24 horas após o infarto agudo do miocárdio (IAM), no entanto, é comum o repouso prolongado no leito em razão do receio de instabilização do paciente. OBJETIVOS: Avaliar as respostas autonômicas e hemodinâmicas de pacientes pós-IAM submetidos ao primeiro dia de protocolo de FTCV fase I, bem como sua segurança. MATERIAIS E MÉTODOS: Foram estudados 51 pacientes com primeiro IAM não complicado, 55 ± 11 anos, 76% homens. Foram submetidos ao primeiro dia do protocolo de FTCV fase I, em média 24 horas pós-IAM. A frequência cardíaca (FC) instantânea e os intervalos R-R do ECG foram captados pelo monitor de FC (Polar®S810i) e a pressão arterial (PA) aferida pelo método auscultatório. A variabilidade da FC foi analisada nos domínios do tempo (RMSSD e RMSM dos iR-R em ms) e da frequência. A densidade espectral de potência foi expressa em unidades absolutas (ms²/Hz) e normalizada (un) para as bandas de baixa (BF) e alta frequência (AF) e pela razão BF/AF. RESULTADOS: O índice RMSSD, a AF e a AFun apresentaram redução na execução dos exercícios em relação ao repouso pré e pós-exercício (p < 0,05), a BFun e a razão BF/AF aumentaram (p < 0,05). A FC e a PA sistólica apresentaram aumento durante a execução dos exercícios em relação ao repouso (p < 0,05). Não foi observado qualquer sinal e/ou sintoma de intolerância ao esforço. CONCLUSÕES: O exercício realizado foi eficaz, pois promoveu alterações hemodinâmicas e na modulação autonômica nesses pacientes, sem ocasionar qualquer intercorrência clínica.

Variabilidade da frequência cardíaca e carga máxima atingida no teste de esforço físico dinâmico em homens idosos

INTRODUÇÃO: Um dos benefícios promovidos pelo exercício físico parece ser a melhora da modulação do sistema nervoso autônomo sobre o coração. No entanto, o papel da atividade física como um fator determinante da variabilidade da frequência cardíaca (VFC) não está bem estabelecido. Desta forma, o objetivo do estudo foi verificar se há correlação entre a frequência cardíaca de repouso e a carga máxima atingida no teste de esforço físico com os índices de VFC em homens idosos. MÉTODOS: Foram estudados 18 homens idosos com idades entre 60 e 70 anos. Foram feitas as seguintes avaliações: a) teste de esforço máximo em cicloergômetro utilizando-se o protocolo de Balke para avaliação da capacidade aeróbia; b) registro da frequência cardíaca (FC) e dos intervalos R-R durante 15 minutos na condição de repouso em decúbito dorsal. Após a coleta, os dados foram analisados no domínio do tempo, calculando-se o índice RMSSD, e no domínio da frequência, calculando-se os índices de baixa frequência (BF), alta frequência (AF) e razão BF/AF. Para verificar se existe associação entre a carga máxima atingida no teste de esforço e os índices de VFC foi aplicado o teste de correlação de Pearson (p < 0,05). RESULTADOS: Características demográficas, antropométricas, fisiológicas e carga máxima atingida no teste ergométrico: idade = 63 ± 3,0 anos; IMC = 24 ± 2kg/m²; FC = 63 ± 9bpm; PAS = 123 ± 19mmHg; PAD = 83 ± 8mmHg; carga máxima = 152 ± 29 watts. Não houve correlação entre os índices de VFC com os valores de FC de repouso e carga máxima atingida no teste ergométrico (p > 0,05). CONCLUSÃO: Os índices de variabilidade da frequência cardíaca temporal e espectrais estudados não são indicadores do nível de capacidade aeróbia de homens idosos avaliados em cicloergômetro.

Análise eletromiográfica dos músculos da coxa no exercício agachamento afundo até a exaustão

O agachamento afundo possui um posicionamento dos membros inferiores diferencial em relação ao agachamento padrão, necessitando de maiores esclarecimentos acerca das participações dos músculos envolvidos. O objetivo foi analisar a atividade eletromiográfica dos músculos vastus lateralis (VL), vastus medialis (VM), bíceps femoris (BF) e semitendinosus (ST) durante a execução do agachamento afundo até à exaustão com o membro inferior posicionado frontalmente e posteriormente. Participaram do estudo nove mulheres ativas com média (DP) de idade de 22 (3,4) anos e massa corporal 60,3 (4,1) kg. O agachamento afundo foi dividido em duas etapas, diferindo apenas o posicionamento do membro inferior dominante (randomizado). Os sinais eletromiográficos foram captados utilizando um eletromiógrafo e analisados os valores "root mean square" (RMS) na fase concêntrica. Os resultados indicaram um aumento significativo do RMS em função do tempo para o membro inferior posicionado frontalmente e posteriormente (p< 0,001). No membro posicionado frontalmente, o aumento do RMS correspondeu a 50% para o VL, 54% para o VM e 48% para o BF. O membro posicionado posteriormente apresentou um aumento de 75% para o VL, 113% para o VM, 62% para o BF e 48% para o ST. O RMS também foi significativamente maior no músculo VM em relação ao ST no membro inferior posicionado anteriormente (p = 0,03) e em relação ao ST e BF no membro inferior posicionado posteriormente (p = 0,02). Não ocorreu interação significativa entre o efeito do tempo e músculo na atividade eletromiográfica. O RMS normalizado não apresentou diferenças estatisticamente significativas no que se refere ao posicionamento do membro inferior dominante. A atividade muscular foi semelhante em ambos os posicionamentos do membro inferior, apresentando maior aumento de ativação dos músculos VL e VM em relação ao BF e ST.