Influence of the temperature and type of salt on the phase equilibrium of peg 1500 + potassium phosphate and peg 1500 + sodium citrate aqueous two-phase systems

The present work analyzed the effect of the temperature and type of salt on the phase equilibrium of aqueous two-phase systems (ATPS) formed by poly (ethylene glycol) (PEG) 1500 + potassium phosphate, from (278.15 to 318.15) K, and PEG 1500 + sodium citrate, from (278.15 to 298.15) K. The rise of the temperature normally increased the slope of the tie line (STL). With respect to the influence of the type of salt, sodium citrate showed better capability to induce phase separation, when compared to potassium phosphate.

Un método reproducible para obtener peg biramificado monofuncional de alta pureza

PEGylation has become a widely applied technique to enhancing in vitro and in vivo stability of therapeutic proteins and to increasing materials biocompatibility. PEG branched structures have proven useful for protein and peptide modification. Furthermore, they may be better than linear structures for many purposes. This paper describes an improved procedure for obtaining 2-arms PEG based on L-lysine. The efficiency of the synthesis was not related to moisture of the raw materials. This procedure does not use hazardous reagents as previous protocols do. It implemented a purification process for obtaining the desired structure with high purity ( > 99%). Finally, the procedure described here allows the obtaining of others PEGylation reagents.

Partition of proteases from Lentinus citrinus DPUA 1535 by the Peg/Phosphate Aqueous Two-Phase System

A full two-level factorial design was employed to study the influence of PEG molar mass (MM PEG), PEG concentration (C PEG) and phosphate concentration (C PHOSPH) on proteases partition by Lentinus citrinus DPUA 1535 in a PEG/phosphate aqueous two-phase system (ATPS). For all ATPS studied, proteases partitioned for the top phase and the best proteases extraction condition was obtained with MM PEG = 6000 g mol-1, C PEG = 17.5% (w/w) and C PHOSPH = 25% (w/w) with (1.1) purification factor and (151%) activity yield. Findings reported here demonstrate a practical strategy that serves as a first step for proteases purification from crude extract by L. citrinus.

Qualidade fisiológica de sementes de algodoeiro submetidas ao condicionamento mátrico e osmótico

Objetivou-se, neste trabalho, avaliar o efeito do condicionamento mátrico e osmótico sobre a qualidade fisiológica de sementes de algodão herbáceo. Foram utilizadas sementes de algodão, deslintadas quimicamente, do cultivar Panton, na proporção de 100 g de sementes para 1000 g da matriz sólida (algalita). Os tratamentos consistiram em cinco teores de água, utilizados para umedecimento da matriz (0,10; 0,25; 0,50; 0,75 e 1,0 mL), durante seis períodos de condicionamento (0; 1; 4; 8; 16 e 32 dias). Quanto ao condicionamento osmótico, foram realizados dois ensaios: sementes submetidas às soluções osmóticas de polietileno glicol (PEG) 6000 (-0,25 MPa) e de nitrato de potássio (0,3 M ou -0,72 Mpa), nos sete tempos de condicionamento osmótico (0; 12; 24; 36; 48; 72 e 96 h), à temperatura de 20 °C. As variáveis analisadas foram porcentagem de germinação e comprimento de plântulas. O delineamento experimental utilizado foi inteiramente ao acaso, com quatro repetições. Observou-se que houve aumento na germinação e no vigor das sementes condicionadas com diferentes níveis de umidade na matriz sólida, em comparação com a testemunha. Constatou-se, também, que a capacidade de revigoramento do tratamento com 0,50 mL de água foi comprovada, à medida que aumentou o tempo de exposição das sementes à matriz sólida; o condicionamento osmótico com PEG 6000 aumentou o vigor das sementes, mas não favoreceu a germinação, e o condicionamento osmótico com nitrato de potássio reduziu a germinação, entretanto, proporcionou aumento no vigor das sementes nas primeiras horas de hidratação.

Tolerância à seca de genótipos de trigo utilizando agentes indutores de estresse no processo de seleção

Com o objetivo de selecionar genótipos de trigo tolerantes à seca, por meio da indução de déficit hídrico por diferentes agentes osmóticos, sementes e embriões imaturos de oito genótipos foram submetidos a diferentes gradientes osmóticos e avaliados quanto ao comprimento da parte aérea e da raiz principal e à produção de biomassa total. As sementes foram submetidas aos agentes polietilenoglicol-6000 e manitol, em sete gradientes osmóticos, e avaliadas após dez dias. Os embriões foram cultivados em meio macrobatata, suplementado com PEG 6000, manitol e maltose nas concentrações de 10, 20 e 30%, além do controle, e avaliados após 30 dias de incubação. O delineamento foi inteiramente casualizado e os dados foram submetidos à análise de regressão, utilizando-se o teste "t", para comparar os valores de Beta, e análise de variância, com teste de médias pelo teste "t". Houve efeito do estresse hídrico em todos os genótipos testados para as plântulas oriundas de sementes, com interação significativa entre agente e cultivar e efeito negativo maior para PEG 6000. Os genótipos BH 1146 e Ocepar 14 apresentaram maior tolerância à seca, redução média no crescimento, na presença dos agentes estressantes, de 58 e 62%, respectivamente. Em contraste, o Aliança foi o mais sensível, com redução média de 92%. Para as plântulas oriundas de embriões imaturos, o genótipo BH 1146 também apresentou relativa tolerância; já o Ocepar 14 e Aliança foram altamente sensíveis. Porém, estes genótipos também apresentaram baixo desempenho no meio sem a presença dos agentes estressantes, podendo o efeito negativo estar condicionado ao cultivo in vitro.

Detection of circulating polysaccharide antigen of Schistosoma mansoni in hamster sera by crossed immunoelectrophoresis

Crossed immunoelectrophoresis (IEC) was used for detection of free and complexed circulating polisaccharide anodic antigen (AgCA) of Schistosoma mansoni in sera of infected hamsters. An attempt was also done to detect AgCA in human sera from patients infected with S. mansoni. The conditions for isolation and detection of complexed AgCA were established. The sensitivity of IEC was increased by incorporation of 2% polyethylene glicol (PEG) to the agarose and by maintaining the system at 4°C during the electrophoretic run. Free AgCA was detected in 12 and the complexed in 30 of the 37 hamsters sera analysed. Correlation between AgCA (free and complexed) and the parasite load was observed. AgCA was not detected, under the experimental conditions used, in human sera from 7 patients in the acute and 23 in the chronic phase of infection.

IMPACT OF THE PEGYLATED-INTERFERON AND RIBAVIRIN THERAPY ON THE TREATMENT-RELATED MORTALITY OF PATIENTS WITH CIRRHOSIS DUE TO HEPATITIS C VIRUS

Although the protease inhibitors have revolutionized the therapy of chronic hepatitis C (CHC), the concomitant use of pegylated-interferon (PEG-IFN) and ribavirin (RBV) is associated to a high rate of adverse effects. In this study, we evaluated the consequences of PEG-IFN and RBV and their relationship with mortality in patients with cirrhosis. METHODS: Medical records of CHC who underwent treatment with PEG-IFN and RBV in a public hospital in Brazil were evaluated. All the patients with cirrhosis were selected, and their clinical and laboratory characteristics, response to treatment, side effects and mortality were evaluated. RESULTS: From the 1,059 patients with CHC, 257 cirrhotic patients were evaluated. Of these, 45 (17.5%) achieved sustained viral response (SVR). Early discontinuation of therapy occurred in 105 (40.8%) patients, of which 39 (15.2%) were due to serious adverse effects. The mortality rate among the 257 cirrhotic patients was 4.3%, occurring in 06/242 (2.4%) of the Child-A, and in 05/15 (33.3%) of the Child-B patients. In conclusion, the treatment of patients with cirrhosis due to HCV with PEG-IFN and RBV shows a low SVR rate and a high mortality, especially in patients with liver dysfunction.

Gender influence on treatment of chronic hepatitis C genotype 1

INTRODUCTION: Although various studies have been published regarding the treatment of chronic hepatitis C (CHC) with peginterferon (Peg-IFN) and ribavirin, little is known regarding the real impact of gender on the characteristics that influence the effectiveness and safety of antiviral treatment for CHC patients. The objective of this study was to evaluate the influence of gender on HCV treatment outcomes. METHODS: A retrospective analytical study was conducted among selected carriers of CHC genotype 1, who were treated with Peg-IFN α-2b at a dose of 1.5 μg/kg or Peg-IFN α-2a at a dose of 180 μg/week plus a ribavirin dose of 1,000-1,250 mg/day, according to weight, between 2001 and 2007. RESULTS: Among 181 patients undergoing treatment, the mean age was 46.4 ± 11.0 years and 46% were women. At baseline, 32% of the patients had advanced fibrosis (F3-F4 Scheuer), and 83% of the subjects had viral load > 400,000 IU/ml, without significant difference between the genders (p = 0.428 and p = 0.452, respectively). When compared with men, women had higher incidence of many adverse events such as anemia (p < 0.001) and higher need for dose reduction, for both Peg-IFN (p = 0.004) and ribavirin (p = 0.006). However, the rate of sustained virological response (SVR) did not differ between the genders: 45% (female) vs 41% (male); p=0.464. CONCLUSIONS: This study suggests that women and men react differently to combined therapy, especially in relation to the incidence of adverse events and the need for dose modification. Nevertheless, these differences do not influence the SVR rate.

Evaluation of the therapeutic response of hepatitis C in coinfected patients (HIV/HCV): a study of cases from a hospital for chronic liver diseases in the Eastern Brazilian Amazon

INTRODUCTION: The aim of this study was to evaluate the therapeutic response of hepatitis C in patients coinfected with human immunodeficiency virus (HIV-1). METHODS: A retrospective study of 20 patients coinfected with HIV-1/HCV who were treated in the outpatient liver clinic at the Sacred House of Mercy Foundation Hospital of Pará (Fundação Santa Casa de Misericórdia do Pará - FSCMPA) from April 2004 to June 2009. Patients were treated with 180µg PEG interferon-α2a in combination with ribavirin (1,000 to 1,250mg/day) for 48 weeks. The end point was the sustained virological response (SVR) rate (HCV RNA negative 24 weeks after completing treatment). RESULTS: The mean age of the patients was 40±9.5 years, of which 89% (n=17) were male, and the HCV genotypes were genotype 1 (55%, n=11/20), genotype 2 (10%, n=2/20) and genotype 3 (35%, n=7/20). The mean CD4+ lymphocyte count was 507.8, and the liver fibrosis stages were (METAVIR) F1 (25%), F2 (55%), F3 (10%) and F4 (10%). The early virological response (EVR) was 60%, the end-of-treatment virological response (EOTVR) was 45% and the SVR was 45%. CONCLUSIONS: The median HCV viral load was high, and in 85% of cases in which highly active antiretroviral therapy (HAART) was used, none of the patients with F3-F4 fibrosis responded to treatment. Of the twenty patients treated, 45% achieved SVR and 45% achieved EOTVR. Studies that include cases from a wider region are needed to better evaluate these findings.

Impact of the IL-18 gene polymorphism in response to antiviral therapy in chronic HCV genotype 4 patients

^a Introduction Interleukin (IL)-18 is a well-known major proinflammatory cytokine with broad biological effects. The major immunomodulatory functions of IL-18 include enhancing T cell and natural killer cell cytotoxicity. Serum levels of this cytokine were shown to increase in chronic hepatitis C patients compared to non-infected healthy people. An association between IL-18 gene promoter polymorphisms and pegylated interferon (PEG-IFN) and ribavirin treatment outcomes has been reported for individuals with chronic hepatitis C virus genotype 1 (HCV-1). In this study, HCV genotype 4 (HCV-4) patients were assessed for IL-18 gene polymorphisms and treatment outcomes or severity of liver disease because data concerning the impact of IL-18 gene polymorphisms on patients with HCV-4 infections are limited. Methods This study included 123 chronic HCV-4 Egyptian patients and 123 apparently healthy volunteer blood donors who served as a control group. HCV genotyping was performed using the line probe assay. IL-18 genotyping was performed using the TaqMan Real-Time PCR method in all 246 patient and control samples. Results In our study, all patients had HCV-4. IL-18 gene single nucleotide polymorphism (SNP) (-607C/A) genotype distributions and allele frequencies did not differ between HCV patients and normal healthy subjects or between patient groups when compared according to the therapeutic response. Moreover, the presence of an IL-18 SNP was not associated with histological disease severity. We conclude that the presence of the IL-18 SNP rs1946518 does not affect the outcome of chronic HCV-4 treatment in Egyptian patients. Conclusions The IL-18 SNP rs1946518 does not affect response to treatment in chronic HCV-4 patients.

Utilização dos recursos naturais por comunidades humanas do Parque Ecoturístico do Guamá, Belém, Pará

O trabalho apresenta dados sobre o uso dos recursos naturais por populações humanas em pequenas comunidades (vilas) localizadas no entorno (n = 6) e no interior (n = 1) do Parque Ecoturístico do Guamá (PEG), visando avaliar os impactos sobre a fauna e flora local, ante a implantação de um projeto de infra-estrutura rodoviária local. A metodologia consistiu da aplicação de dois modelos de questionários. Um direcionado a pessoas-chave, como: moradores mais antigos, presidentes de associações, agentes de saúde, etc, e o outro aplicado aleatoriamente aos adultos locais. Buscou-se atingir pelo menos 50% das famílias das pequenas localidades (<10 famílias) e 10-15% no caso de comunidades maiores ( >11 famílias). Em todos os casos é realizado o extrativismo da fauna e flora, principalmente com o objetivo de subsistência. As espécies mais caçadas foram Agouti paca,Dasyprocta agouti,Nasua nasua, Euphractus sexcintus e Hydrochaeris hydrochaeris. A complementação da renda familiar através do comércio de caça sob encomenda é realizada em todas as comunidades estudadas. Frutíferas como Euterpe oleracea Mart., Musa spp e Theobroma cacao foram encontradas em 100% das comunidades (n=7). Plantas medicinais são rotineiramente cultivadas e empregadas. As espécies madeireiras exploradas são utilizadas com o objetivo de produção de carvão, construção de casas e de meios de transporte, predominando o Inga edulis, Virola surinamensis (Rol.) Warb e Simaruba amara (Aubl.). O trabalho discute os resultados sob o aspecto da implantação do projeto viário, oferecendo recomendações para a minimização dos impactos do mesmo sobre a fauna, a flora e o modo tradicional de vida das comunidades.

Additives and Protein-DNA Combinations Modulate the Humoral Immune Response Elicited by a Hepatitis C Virus Core-encoding Plasmid in Mice

Humoral and cellular immune responses are currently induced against hepatitis C virus (HCV) core following vaccination with core-encoding plasmids. However, the anti-core antibody response is frequently weak or transient. In this paper, we evaluated the effect of different additives and DNA-protein combinations on the anti-core antibody response. BALB/c mice were intramuscularly injected with an expression plasmid (pIDKCo), encoding a C-terminal truncated variant of the HCV core protein, alone or combined with CaCl2, PEG 6000, Freund's adjuvant, sonicated calf thymus DNA and a recombinant core protein (Co.120). Mixture of pIDKCo with PEG 6000 and Freund's adjuvant accelerated the development of the anti-core Ab response. Combination with PEG 6000 also induced a bias to IgG2a subclass predominance among anti-core antibodies. The kinetics, IgG2a/IgG1 ratio and epitope specificity of the anti-core antibody response elicited by Co.120 alone or combined with pIDKCo was different regarding that induced by the pIDKCo alone. Our data indicate that the antibody response induced following DNA immunization can be modified by formulation strategies.

Response to treatment in Brazilian patients with chronic hepatitis C is associated with a single-nucleotide polymorphism near the interleukin-28B gene

A single-nucleotide polymorphism (SNP) upstream of interleukin (IL)28B was recently identified as an important predictor of the outcome of chronic hepatitis C patients treated with pegylated interferon plus ribavirin (PEG-IFN/RBV). The aim of this study was to investigate the association between the IL28B gene polymorphism (rs12979860) and virological response in chronic hepatitis C patients. Brazilian patients (n = 263) who were infected with hepatitis C virus (HCV) genotype 1 and were receiving PEG-IFN/RBV were genotyped. Early virological response (EVR) (12 weeks), end-of-treatment response (EOTR) (48 weeks), sustained virological response (SVR) (72 weeks) and relapse were evaluated using conventional and quantitative polymerase chain reaction (PCR) assays. The frequency of the C allele in the population was 39%. Overall, 43% of patients experienced SVR. The IL28B CC genotype was significantly associated with higher treatment response rates and a lower relapse rate compared to the other genotypes [84% vs. 58% EVR, 92% vs. 63% EOTR, 76% vs. 38% SVR and 17% vs. 40% relapse rate in CC vs. other genotypes (CT and TT), respectively]. Thus, the IL28B genotype appears to be a strong predictor of SVR following PEG-IFN/RBV therapy in treatment-naïve Brazilian patients infected with HCV genotype 1. This study, together with similar research examining other SNPs, should help to define adequate protocols for the treatment of patients infected with HCV genotype 1, especially those with a poor prognosis.

Health-Related Quality of Life in patients with hepatitis C in double and triple therapy

Abstract OBJECTIVE Comparing Health-Related Quality of Life (HRQoL) scores in patients with chronic hepatitis C undergoing double and triple antiviral therapy and analyzing possible factors related to HRQoL. METHOD HRQoL was assessed using the Short Form 36 and Chronic Liver Disease Questionnaire, which were applied at baseline and at weeks 4, 12 and 16 of treatment to 32 patients divided into two groups: double therapy with pegylated interferon (IFN-PEG) and ribavirin, and triple therapy with PEG-IFN, ribavirin and telaprevir. RESULTS The reduction of HRQoL was greater in patients receiving triple therapy compared to those treated with two drugs, the most critical time is at 12 weeks in both groups. After removal of telaprevir, the triple therapy group significantly improved their HRQoL scores. Anxiety and depression before treatment, employment status and race are significantly related to diminished HRQoL. CONCLUSION Patients undergoing double and triple therapy have diminished HRQoL indexes, but the addition of telaprevir chooses a more significant decrease.

Avaliação da qualidade fisiológica de sementes de soja, após o processo de hidratação-desidratação e envelhecimento acelerado

Foi conduzido um experimento com o objetivo de avaliar a qualidade fisiológica de sementes de soja (Glycine max (L.) Merrill) , envolvendo as variedades UFV-10, IAC-8 e Doko RC, submetidas ao processo de hidratação-desidratação, seguido por uma condição de estresse induzida pelo teste de envelhecimento acelerado. As sementes foram pré-condicionadas em PEG 6000, com potencial osmótico de -0,8 MPa, a uma temperatura de 20ºC, por quatro dias. Em seguida, as sementes foram novamente desidratadas, até atingir o conteúdo de umidade inicial, sendo então submetidas ao teste de envelhecimento acelerado pelos seguintes períodos: 0, 24, 48 e 72 horas. As sementes pré-condicionadas foram comparadas com duas testemunhas: sementes não-tratadas e sementes embebidas em água. Após cada período de envelhecimento, as sementes foram avaliadas pelo teste padrão de germinação, comprimento de plântula e biomassa seca das plântulas. Em casa de vegetação, determinaram-se o índice de velocidade de emergência e a emergência final em substrato de areia. O tratamento de hidratação-desidratação em solução de PEG 6000 melhorou o desempenho das sementes em termos de germinação e vigor. O aumento nos períodos de envelhecimento acelerado influenciou negativamente o desempenho das sementes, em todos os tratamentos avaliados. Contudo, o tratamento com PEG 6000 foi satisfatório em manter a qualidade das sementes. A embebição das sementes de soja em água desmineralizada prejudicou a qualidade fisiológica. As variedades de soja mostraram resposta diferenciada aos tratamentos de hidratação-desidratação e aos períodos de envelhecimento acelerado.