Comparison of risk factors among blood donors, volunteers and replacement individuals, infected or not by hepatitis C virus

Hepatitis C is transmitted primarily parenterally by contaminated blood and is often associated with: intravenous drug abuse, invasive procedures, blood transfusions, acupuncture, tattooing, and alcohol and tobacco use. This study aimed to quantify and evaluate the risk factors among blood donors, volunteer blood donors and replacement individuals, infected or not by the C virus. The main transmission routes of C virus were identified in 55 men and 25 women (GI) monitored by the Ambulatory Unit of the Department of Tropical Diseases, Botucatu Medical School, and 24 men and 26 women (GII), all active blood donors at the Bauru State Hospital Transfusional Agency. Both groups were similar in: tobacco and alcohol consumption, sexual behavior, tattooing and illicit drug use. The duration of alcohol and tobacco consumption and blood transfusions in GI were longer, whereas the option for steady partners, condom use, disposable materials and piercings were predominant in GII. In conclusion, the risk factors for hepatitis C demonstrate the necessity of health policies that act on the primary and secondary prevention levels (respectively, reduction of infection incidence and hepatopathy risk)

Assessment of cytokine values in serum by RT-PCR in HIV-1 infected individuals with and without highly active anti-retroviral therapy (HAART)

A cross-sectional study was performed on HIV-1 infected individuals with or without antiretroviral treatment (ARV) in the AIDS Day Hospital, Botucatu Medical School, UNESP. Between August 2004 and October 2005, 73 HIV-1 infected individuals were divided into three groups: infected individuals with or without AIDS who had never received ARV (G1 = 15); patients on HAART that had had plasma HIV-1 RNA viral load (VL) equal to or greater than 50 copies/mL (G2 = 27); and patients on HAART with undetectable VL for at least the past six months (G3 = 31). There was also an additional group that comprised blood donors without any sign of the disease and with negative HIV serum tests (G4 = 20), which was the control group. Serum cytokine levels (values in pg/mL) were measured by enzyme-linked immunosorbent assay (ELISA) and specific mRNA expression by reverse transcription polymerase chain reaction (RT-PCR). Both techniques were performed on the four groups for TNF-á, IL-2, INF-ã, IL-4 and IL-10. All patients were submitted to VL determination and CD4+ and CD8+T lymphocyte counts. The analysis of the results revealed a significant comparison among groups for both methods and an association between the latter (> 80% r² > 0.80). There was only one exception, in control individuals for IL-2 by ELISA. The cytokine profiles, in both methods, for the three patient groups, were mature Th-0. The behaviors of IL-2 and INF-ã required emphasis due to consequent expression of dominant Th profile. Both methods showed low IL-2 and high mean values of INF-ã in the three groups. Several authors have recently drawn attention to the substantial apoptosis of infected and non-infected CD4+T cells, mainly during primary infection, persisting only in those with INF-ã phenotype producer and not IL-2. HIV infected individuals submitted to HAART are expected to produce IL-2 in an attempt to present Th-1 profile, but in most cases this did not occur.(AU)

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 µg/g in the liver and 51.2 µg/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.(AU)

Cytomegalovirus and Toxoplasma gondii seroprevalence in a Brazilian liver transplant waiting

Cytomegalovirus (CMV) disease is a major cause of morbidity and mortality in solid organ transplantation. Disseminated toxoplasmosis after liver transplantation is a rare but fatal event. Serologic screening of the donor and the recipient is essential to prophylactic management, early diagnosis and therapeutic strategies to minimize the consequences of these infections. The aim of the present study was to determine the seroprevalence of CMV and Toxoplasma gondii (TG) in a Brazilian liver transplant waiting list (LTWL). Serological data were collected from 44 candidates on the LTWL between May 2003 and November 2004. Serological investigation of antibodies IgM and IgG against CMV (anti-CMV) and TG (anti-T. gondii) was performed using fluorometry commercial kits. IgG anti-CMV was positive in 37 patients (94.9 percent) out of 39 available results. There were not IgM anti-CMV positive results. Out of 36 analyzed patients, 22 (61.1 percent) presented positive IgG anti-T. gondii and none had positive IgM anti-T. gondii. The high CMV seroprevalence among our LTWL reinforces the need for appropriate protocols to avoid related complications, like reactivation and superinfection by CMV. Environmental and drug prophylactic strategies against primary infection and reactivation, as well as early diagnosis and treatment of toxoplasmosis complications, are essential for the good outcome of transplant patients.

Laboratory evaluation of young ovines inoculated with natural or 60co-irradiated Crotalus durissus terrificus venom during hyperimmunization process

Laboratory profile of young ovines was studied in order to evaluate and compare their antiserum production from natural and Cobalt-60 irradiated Crotalus durissus terrificus (C.d.t.) venoms. The parameters analyzed included complete blood count, and urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin serum measurements. Three groups of six animals each were used. Group 1 (G1) received natural C.d.t. venom; Group 2 (G2) received irradiated C.d.t. venom; and Group 3 (G3) was used as control and did not receive venom, only adjuvants, using seven venom inoculations. During the experimental period, animals were fortnightly weighed. According to clinical and weight evaluation, sheep in post-weaning phase showed no changes in their physiological profiles but had excellent weight gain. The parameters analyzed were not statistically different (p<5 percent) among the groups tested. The hyperimmunization process was successfully accomplished with the production of specific antibodies against Crotalus durissus terrificus venom. Results bring a new possibility of utilizing ovines in the commercial production of anticrotalic serum, which may be used to treat human and animal envenomation. Its production cost may be reduced by subsequent use of hyperimmunized sheep for human consumption.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD45RA and CD45RO cell count; determination of IL-2, IFN-gamma, IL-4, IL-10 and TNF-alpha serum levels; CD4+ T and CD8+ T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD45RA (G1<[G3=GC-2]; p<0.05) and CD45RO (G1[G2=G3]; p<0.001), TNF-alpha serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-gamma ([G1=GC-1]<[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD8+ T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-gamma), CD45RA and CD45RO cells, and CD4+ T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-alpha, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-gamma, and VL, followed by CD45RA and CD45RO cells, and IL-10...

Immunological assessment of mice hyperimmunized with native and cobalt-60-irradiated Bothrops venoms

ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated (60Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom

Trypanosomatids in dogs belonging to individuals with chronic chagas'disease living in Botucatu town and surrounding region, São Paulo State, Brazil

Among domestic animals, dogs are considered to be the major reservoirs of trypanosomatids and, due to their proximity to man, the presence of these parasites in dogs is an alert to actions aiming at triatomine control. Fifty dogs (26 males and 24 females), aged from 2 months to 15 years, belonging to 30 chronic Chagas’ disease individuals from 15 different municipalities in the western region of São Paulo State, Brazil, were subjected to blood collection for the following tests: artificial xenodiagnosis, blood culture, and Polymerase Chain Reaction (PCR). Forty-three (86%) out of 50 dogs were positive to at least one of the tests performed; 34 (68%) were positive to xenodiagnosis, 30 (60%) to blood culture, and 25 (50%) to PCR for T. cruzi and/or T. rangeli. Although triatomines were not detected during the intra and peridomiciliary inspections in the dog owners’ residences, the results obtained demonstrate that there is a transmission cycle whereby triatomine vector may be participating in the infection epidemiological chain

Immunization with native and cobalt 60-irradiated Crotalus durissus terrificus venom in swiss mice: assessment of the neutralizing potency of antisera

ELISA was used to evaluate, follow, and compare the humoral immune response of Swiss mice during hyperimmunization with natural and Cobalt 60-irradiated (60Co) Crotalus durissus terrificus venom. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by “in vivo” challenge and the side effects were assessed. The animals immunization with one LD50 of the venom was on days one, 15, 21, 30, and 45, when blood samples were collected; the challenges occurred on the 60th day. Results showed that ELISA was efficient in evaluating, following, and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co irradiation. Serum produced from Crotalus durissus terrificus irradiated venom showed higher potency and neutralization capacity than that from natural venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom

Correlation between cytokine serum levels, number of CD4+ T cells/mm³ and viral load in HIV-1 infected individuals with or without antiretroviral therapy

Seventy-nine HIV-1 infected patients were studied in three groups: Group G1 - 11 patients with no antiretroviral therapy; G2 - 40 patients undergoing antiretroviral therapy, 33 with only two nucleoside reverse transcriptase inhibitors (NRTI), and seven with two NRTI and one protease inhibitor (PI), all with viral load (VL) equal or higher than 80 copies of plasma RNA/ml; Group G3 - 28 patients, 23 on highly active antiretroviral therapy (HAART), 18 with two NRTI and one PI, and five with two NRTI and one non-nucleoside reverse transcriptase inhibitor (NNRTI), the remaining five with combination of two NRTI. All G3 patients had undetectable viral load for at least the past six months. The control group (Gc) included 20 normal blood donors without clinical complaints or signs of disease and negative for anti-HIV-1/2 antibodies. Serum cytokine levels pg/ml (TNF-alpha, INF-gamma, IL-2, IL-4, and IL-10) were determined in all patients including controls. CD4+ T and CD8+ T lymphocyte counts were made in the 79 patients by flow cytometry; VL determination was by NASBA technology. Analysis of results showed that the number of CD4+ T and CD8+ T lymphocytes were higher in G2 than G1, while VL was 0.5 log lower. G3 patients had similar lymphocyte values to G2, however they were chosen for G3 because their VL was undetectable, different by 4.0 log to G2. These results show the effect of antiretroviral treatment in G2 and G3 patients with better performance in the latter. Statistical difference was seen between the three groups and controls for serum cytokine behavior: TNF-alpha [H=48.323; p<0.001;(G1=G2=G3)>Gc]; INF-gamma[H=28.992; p<0.001; (G1=G2=G3)>Gc]; IL-4[H=48.323; p<0.001; (G1=G2=G3)>Gc]; IL-10[H=47.256; p<0.001; (G1=G2=G3)>Gc. There was no statistical difference in IL-2 values between all groups (H=6.071; p>0.10; G1=G2=G3=Gc). In absolute values however, G3 showed slightly lower TNF-alpha, IL-4, and IL-10, and higher INF-gamma and IL-2, to G1 and G2. This suggests a better performance in G3 patients, especially in IL-2 behavior. For cytokine profile, the three groups showed mature Th0 subset. In G1 72.73 percent were mature Th0, and 27.27 percent Th2; G2, 72.50 percent mature Th0, and 27.50 percent Th2; and G3, 89.29 percent mature Th0, and 10.71 percent Th2. There was no statistical difference between groups (chi(2)2=3.014; p>0.10; G1=G2=G3). Statistical difference was seen between G2 and G3 for antiretroviral regimes used...

Down-modulation of lymphoproliferation and interferon-gamma production by beta-glucan derived from Saccharomyces cerevisiae

Beta-glucan, one of the major cell wall components of Saccharomyces cerevisiae, has been found to enhance immune functions. This study investigated in vivo and in vitro effects of beta-glucan on lymphoproliferation and interferon-gamma (IFN-gamma) production by splenic cells from C57BL/6 female mice. All experiments were performed with particulate beta-glucan derived from S. cerevisiae. Data demonstrated that both, i.p administration of particulate beta-glucan (20 or 100 µg/animal) and in vitro stimulation of splenic cells (20 or 100 µg/ml of culture) decreased lymphoproliferation and IFN-gamma production induced by concanavalin A. These results suggest that beta-glucan can trigger a down-modulatory effect regulating a deleterious immune system hyperactivity in the presence of a strong stimulus.

Acute-phase reactions, including cytokines, in patients bitten by Bothrops and Crotalus snakes in Brazil

Thirty-one patients bitten by venomous snakes in Botucatu area (State of Säo Paulo - Brazil), sixteen by Bothrops spp. and fifteen by Crotalus durissus terrificus, were studied. The group comprised twenty-nine males and two females, ranging from fourteen to sixty-three years of age (mean 33 ñ 15.Leukocytosis with neutrophilia and lymphopenia, increase of mucoproteins and C reactive protein, decrease of total serum protein and albumin, were observed on the first day after the accident. In addition, increased serum levels of the cytokines IL-6 and IL-8, but not of IL-1 beta and TNF-alpha, were observed. The alterations were generally more intense in patients bitten by Crotalus durissus terrificus than by Bothrops spp. It is concluded that these snakebite envenomations closely resemble an acute trauma, inducing a typical acute-phase response.

Avaliacao do tratamento da paracoccidioidomicose com o ketoconazol. / Evaluation of the treatment of paracoccidioidomycosis with ketoconazole

Foram tratados 12 doentes atendidos na Disciplina de Molestias Infecciosas e Parasitarias da Faculdade de Medicina de Botucatu com diagnostico etiologico de paracoccidioidomicose que apresentavam lesoes organicas multiplas e evolucao prolongada.O tratamento foi realizado por 18 meses, com o Ketoconazol, pela via oral, nas doses diarias de 400 mg no primeiro mes e de 200 mg nos meses seguintes. Todos os doentes foram acompanhados durante o tratamento e, em media 4 meses e meio apos o mesmo, clinica, radiologica e sorologicamente pelas reacoes de imunofluorescencia indireta, precipitinas e imunodifusao em gel.A competencia imunitaria foi avaliada em todos os doentes antes do tratamento e repetida em quatro, no final do mesmo. Os resultados mostraram que houve recaida em 5 doentes. A droga foi bem tolerada e a imunodifusao em gel e a hemossedimentacao foram as provas que mostraram maior paralelismo com a evolucao clinica

Malaria at Humaita County, Amazonas State, Brazil. Some aspects.

Dede 1976, varios aspectos de malaria tem sido estudados no Municipio de Humaita, Estado do Amazonas, por pesquisadores da Faculdade de Medicina e do Instituto Basico de Biologia Medica e Agricola do Campus da Universidade Estadual Paulista "Julio de Mesquita Filho". Neste sentido, foram estudados aspectos epidemiologicos, clinicos, sorologicos, bioquimicos e parasitologicos. O comportamento da populacao e dos doentes tambem foi considerado pela determinacao dos sistemas de grupamento sanguineo e pela investigacao de alguns fatores de resistencia natural (sistema Duffy e Hemoglobina S)

Malaria no municipio de Humaita, Estado do Amazonas - XIII - Uso da clindamicina no tratamento de doentes com infeccao causada pelo Plasmodium falciparum. / Malaria in the municipality of Humaita, State of Amazonas.XIII - Use of clindamycin in the treatment of patients with infection caused by Plasmodium falciparum

Em 1981, 52 doentes com malaria causada por pelo Plasmodium falciparum foram tratados com 20 mg/kg/dia de clindamicina, pelas vias oral e/ou intra-venosa, durante 5 a 7 dias. Os doentes eram examinados diariamente no 14o. e 21o. dias, a contar do inicio do tratamento, eram submetidos a exame clinico e parasitologico de sangue.Cinco (9,7%) tinham malaria grave e 16 (30,8%) tinham sido previamente tratados com cloroquina, pirimetamina e sulfas de acao ultra-lenta, sem sucesso. Os resultados mostraram que todos os doentes se curaram. Os parasitologicos de sangue tornaram-se negativos, em geral em torno do 5o.dia de tratamento. A clindamicina parece ser eficiente, principalmente nos casos graves de malaria causada por Plasmodium falciparum resistente

Malaria no municipio de Humaita, Estado do Amazonas. XIV - Inquerito coprologico em habitantes da regiao em relacao as taxas de hemoglobina. / Malaria in the municipality of Humaita, State of Amazonas. XIV - Coprological survey in inhabitants of the region in relation to hemoglobin levels

Em outubro de 1981, foram estudados 213 individuos do Municipio de Humaita, Estado do Amazonas, dos quais 91 eram habitantes de localidades situadas ao longo da calha do Rio Madeira e 122 do Bairro da Olaria, na zona urbana. Todos foram submetidos a inquerito clinico, epidemiologico e parasitologico de fezes pelas tecnicas de BAERMANN, FAUST & col. e HOFFMAN, PONS & JANER. De 65 habitantes da zona urbana e de 43 do Rio Madeira, foi feita determinacao da hemoglobina. De 25 habitantes da zona urbana e de 16 Rio Madeira, foi feita determinacao hemoglobina A2 pela cromatografia em microcoluna de D.E.A.E. celulose Os resultados revelaram que houve maior proporcao de infestacao parasitaria unica ou multipla entre os habitantes do Rio Madeira. Estes achados pode ser explicado pela diferenca das condicoes de higiene locais, pois, os habitantes do Rio Madeira nao dispoe de qualquer elemento de saneamento basico. Essas diferencas, contudo, nao influiram nos niveis de hemoglobina e de hemoglobina A2 que se mostraram semelhantes nos dois grupos

Malaria no municipio de Humaita, Estado do Amazonas. XV - Inquerito parasitologico em dias consecutivos nos habitantes da regiao. / Malaria in the municipality of Humaita, State of Amazonas. XV - Parasitological survey in consecutive days in inhabitants of the region

Em agosto de 1979, foram estudados 293 individuos do Municipio de Humaita, dos quais 105 eram habitantes dos povoados situados as margens do Rio Madeira, 72 eram indios da Tribo Tenhairim e 116 residentes da zona urbana de Humaita. Cada individuo foi examinado de uma a quatro vezes em dias consecutivos. De todos foi feito esfregaco com sangue obtido pela digito-puncao, fixado pelo metanol, corado pelo Giemsa e lido em microscopio de imersao (1.000 X). Os resultados obtidos revelaram apenas 6 casos positivos (2,04%), sendo 3 habitantes do Rio Madeira e 3 indios. Este metodo, alem de trabalhoso, nao trouxe nenhuma contribuicao para o aumento de taxa de positividade em inquerito parasitologico realizado em area endemica

Malaria no municipio de Humaita, Estado do Amazonas. II - Alguns aspectos epidemiologicos comparativos entre 1976 e 1979. / Malaria in the municipality of Humaita, State of Amazonas. II - Some comparative epidemiological aspects of 1976 and 1979

Em 1976 foram estudados 409 e em 1979 foram observados 330 individuos, habitantes das diversas regioes do Municipio de Humaita em povoados situados ao longo das estradas, em localidades na calha do Rio Madeira e na zona urbana.Foram feitas observacoes clinicas completas de todos os individuos estudados, incluindo os antecedentes epidemiologicos. O estudo epidemiologico compreendeu os seguintes elementos: historia pregressa de surtos de malaria, presenca de febre, esplenomegalia e indice esplenico. No seu conjunto os resultados indicam ser a malaria mais frequente, grave e persistente nas estradas, ao contrario das localidades situadas ao longo do Rio Madeira onde ela mostrou tendencia a diminuir no periodo de 1976 a 1979 e sugerem ainda sua ausencia na zona urbana