Cytokines profile and clinical-metabolic alterations in HIV-1-infected individuals with and without lypodistrophy

Highly active antiretroviral therapy (HAART) has lead to a dramatic decrease in the morbidity of patients infected with HIV. However, metabolic side effects, including lipodystrophy and dyslipidemia, have been reported in patients treated with antiretroviral therapy (HAART). The aim of this study was to analyze the clinical and metabolic alterations and the cytokines TNF-α, IFN-γ, IL-2, IL-10 and TNF-II receptors profile in the serum of treated HIV-1-infected individuals with or without lipodystrophy. Eighty-four adult patients were analyzed, 42 females and 42 males, their mean age was 37 years old, and they received HAART for at least 15 months. These patients were ambulatory outpatients from the Infectious and Parasitary Disease Area of Botucatu School of Medicine, UNESP. Subsequently the individuals were distributed into 2 groups, G1: 42 HIV-infected individuals with lipodystrophy, and G2: 42 HIV-infected individuals without lipodystrophy. Among the antiretrovirals used, stavudine was more associated to the lipodystrophy group and zidovudine to the group without lipodystrophy. CD4, CD8, viral load, glucose, albumin, and the circulating lipid did not present any difference in the group comparison, except for triglyceride that was elevated in the lipodystrophy group and HDL which was present in low concentration in more patients of G1. The cytokines TNF-α, TNF-RII, and IL-10 profile presented high levels in the lipodystrophy group; also it was positively correlated with this group. On the other hand, IL-2 and IFN-γ presented low levels in this group. High levels of TNF-α and its receptor seem to be associated to the development of lipodystrophy in patients receiving HAART.

Use of skin graft punch graft type for the healing of leg ulcers in treated Hansen's disease patients

Hansen's disease is an infectious illness caused by Mycobacterium leprae. It affects preferentially the skin and the peripheral nervous system leading to incapacities, such as leg ulcers, which happens due to the direct action of the bacillus on the organs or its indirect action on the peripheral nervous system. Leg ulcers can occur by two physiopathologic processes. There are many treatments for general leg ulcers, which include the ones caused by Hansen's disease sequels. Among them, surgical treatment shows to be effective when using skin graft, which can be performed by several techniques. Considering the low number of techniques known for treating leg ulcers in Hansen's disease sequels, the aims of this work were to standardize alternative techniques, to detect the main bacteria found in ulcer secretion cultures, to analyze the patients profile and the ulcers, to describe the histophatologies found, and to correlate these data with those of literature from all over the world. Skin graft punch type was carried out and analyzed; males had a mean age of 59.4 years old and females, 54.2 years old. Patients were 73.6% male and 26.3% female. Lepromatous type was present in 89.4% patients and tuberculoid type was seen in 10.5% of them. Associated systemic diseases were observed in 26.3% patients. Mean time of ulcers evolution was 11.6 years in male and 12.8 years in women. The average diameter of ulcers in the pre-treatment period was 8.5 X 9.5 cm in male and 10.2 X 6.8 cm in women. After the graft, their average diameters were 3.2 X 2.7 cm in male and 5.1 X 5.6 cm in women. Statistical analysis showed that there was no significant correlation between the ulcer diameter and its reduction or not in the post-surgery period (p=0.269732). The mean age of patients whose ulcers diameter did not change or reduced by only 20% was 63.5 years. Using the Spearman's coefficient, it was possible to observe that there was no significant correlation between the patients' age and the ulcers diameter reduction after the skin graft (p=0.222531). Evolution time of ulcers that did not present any satisfactory result in the post-surgery period was 12.1 years. The Spearman's coefficient showed that there was no significant correlation between the ulcers evolution time and the ulcers diameter reduction in the post-surgery period (p=0.191655). Cultures presented 50% of cases with Pseudomonas aeruginosa. Statistical analysis showed there is no correlation between the bacterial types found and the ulcer evolution in the post-surgery period (p=0.697531). The average of the ulcers diameter reduction was 42.4%, and in 26.3% of the patients the lesions disappeared after the skin graft.

Antibody and cytokine serum levels in patients subjected to anti-rabies prophylaxis with serum-vaccination

Rabies is considered a fatal disease once clinical symptoms have developed. The aim of this study was to evaluate epidemiological aspects and immune response in patients attacked by domestic and wild animals and subjected to post-exposure rabies treatment with equine serum and associated vaccine. Thirty-three patients were evaluated; they were between 13 and 65 years old, 75.8% were male and 24.2% female, and from the Botucatu neighborhood. Twenty healthy control individuals with the same age range were also studied. Specific antibodies to equine immunoglobulins and IFN-γ, IL-2, IL-4, and IL-10 production were evaluated by ELISA. IgM, IgE, IgG and subclasses, and rabies virus antibodies serum levels were determined by nephelometry and seroneutralization methods, respectively. No anaphylactic or serum sickness allergic reactions were observed in patients after treatment. Anti-equine IgG levels were significantly higher than those of IgM after 14 and 28 days of treatment. Protective antibodies to rabies virus > 0.5 UI/ml were detected in 84.6% and 75% of patients at days 14 and 28, respectively. IFN-γ, IL-2 and IL-10 levels in patients before and 48h after treatment were significantly higher than in controls suggesting that both Th1 and Th2 cells were activated in the patients. Serum IgM levels were higher at day 14, and IgG 2 and IgE levels were higher at day 28 of treatment. These results suggest that post-exposure rabies treatment in humans induces significant alterations in patient immune response characterized by increased levels of cytokines, serum levels of specific rabies virus antibodies, and the equine serum components employed in the treatment.

Interval between first dose and booster affected antibody production in cattle vaccinated against rabies

In this study, we compared the levels of neutralizing antibodies induced by inactivated rabies vaccine in cattle by using three alternative immunization procedures. Forty-five bovines (breed nelore) were then organized in three groups (A, B and C, with 15 animals/group). Group A received only one vaccine dose at day zero and Group B received the first dose at day zero and then another dose at day 30 (early booster). Group C was also immunized with two doses; however, the booster was postponed until day 180 after the first dose (delayed booster). Blood samples were withdrawn at days zero (before the first dose) and 30, 210, 390, and 540 after the beginning of immunization and the antibody titers were evaluated by mouse neutralization test. The protocol used to immunize Group C (booster at day 180) was clearly more efficient. In this group, antibody levels were higher and also remained higher for longer periods in comparison with the other two groups. These results show that booster timing significantly affected antibody levels. Therefore, programs addressed to control this disease in cattle should consider not only the use of a booster but also its administration time.

Evaluation of the antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolated from human infections

Multiresistant Staphylococcus aureus constitutes an important public health problem, especially in view of its possible spread in nosocomial environments. In the present work, we analyzed the susceptibility profile of 80 S. aureus stains from human infections resistant to at least 10 drugs. For this study, the techniques used were the disk method and minimum inhibitory concentration (MIC) of the following drugs: cefuroxime, ciprofloxacin, clindamycin, erythromycin, gentamycin, imipenem, oxacillin, rifampicin, tetracycline and vancomycin, according the criteria of the National Committee for Clinical Laboratory Standards (NCCLS). Methicillin was included in the antibiogram as a marker, which is usually used in drugs selection for the treatment of staphylococcal infections. Results indicated that the most effective drug was vancomycin. For the other 10 drugs, the percentage of resistant strains ranged from 85% to 93.75%. In relation to the MICs, it was observed that vancomycin (MIC 90% = 0.615ug/ml) was the most effective drug; followed by rifampicin (MIC 90% = 2.6ug/ml) and ciprofloxacin (MIC 90% = 26.6ug/ml). The drugs that showed the least effective activity were cefuroxime, clindamycin, erythromycin, gentamycin, and oxacillin. On the other hand, observation of β-lactamase production revealed that most of the methicillin-resistant strains produced β-lactamase (83.7%), potentiating the risks of nosocomial infections. In general, vancomycin still continues to be one of the most effective drugs for staphylococcal infections therapy.

Bacterial contamination of the catfish (Cathorops agassizii - Valenciennes, 1839 and Genidens genidens - Agassiz, 1829) sting

This work aimed to study the bacterial contamination in stings of the catfish Genidens genidens and Cathorops agassizii found in the São Vicente estuarine system (São Paulo State, Brazil). For bacteriological analyses, we used fish samples distributed into a group of 50 specimens (25 C. agassizii and 25 G. genidens) and a group of 14 specimens (7 C. agassizii and 7 G. genidens). Results showed contamination of 13 different bacterial species of Enterobacteriaceae, being Klebsiella pneumoniae the most frequent bacteria (26.80%) followed by Enterobacter sp and Escherichia coli (16.27%), and Serratia marcescens, Serratia sp. and Proteus mirabilis (1.16%). Gram-positive bacteria as well as fungi were not detected in the samples. According to the Gram-negative species characterized and with regard to the environmental conditions, it can also be considered that accidents with these catfish stings may develop significant acute secondary infections in humans.

Risk of peritonitis during peritoneal dialysis in carriers of Staphylococcus aureus and coagulase-negative staphylococci

The presence of Staphylococcus aureus in the nasal cavities and pericatheter skin of peritoneal dialysis patients put them at high risk of developing peritonitis. However, it is not clear whether the presence of coagulase-negative staphylococci (CNS) in the nasal passages and skin of patients is related to subsequent occurrence of peritoneal infection. The aim of the present study was to verify the relationship between endogenous sources of S. aureus and CNS and occurrence of peritonitis in patients undergoing peritoneal dialysis. Thirty-two patients on peritoneal hemodialysis were observed for 18 months. Staphylococcus species present in their nasal passage, pericatheter skin and peritoneal effluent were identified and compared based on drug susceptibility tests and dendrograms, which were drawn to better visualize the similarity among strains from extraperitoneal sites as well as their involvement in the causes of infection. Out of 288 Staphylococcus strains isolated, 155 (53.8%) were detected in the nasal cavity, 122 (42.4%) on the skin, and 11 (3.8%) in the peritoneal effluent of patients who developed peritonitis during the study. The most frequent Staphylococcus species were CNS (78.1%), compared with S. aureus (21.9%). Among CNS, S. epidermidis was predominant (64.4%), followed by S. warneri (15.1%), S. haemolyticus (10.7%), and other species (9.8%). Seven (64%) out of 11 cases of peritonitis analyzed presented similar strains. The same strain was isolated from different sites in two (66%) out of three S. aureus infection cases. In the six cases of S. epidermidis peritonitis, the species that caused infection was also found in the normal flora. From these, two cases (33%) presented highly similar strains and in three cases (50%), it was difficult to group strains as to similarity. Patients colonized with multidrug-resistant S. epidermidis strains were more predisposed to infection. Results demonstrated that an endogenous source of S. epidermidis could cause peritonitis in peritoneal dialysis patients, similarly to what has been observed with S. aureus.

Cytokine and antibody production during murine leptospirosis

The aim of the present study was to investigate the kinetics of humoral and cellular responses during leptospirosis. We observed that the presence of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) was associated with antibody production and bacterial recovery, and the compromising of both TNF-α and IL-6 in the immunopathogenesis of leptospirosis during an experimental infection of BALB/c mice inoculated with Leptospira interrogans serovar Canicola was verified. Results showed higher levels of TNF-α and IL-6 in the initial phase of infection, in which the greatest bacterial clearance was observed. However, when the bacterial recovery was compared with the kinetics of the production of antibodies, the results revealed a kinetics proportionally inverted to antibody production. This fact may be related to some inhibitory factor which could be responsible for the selective suppression of the cellular immune response. We concluded that during leptospirosis there was a greater mobilization of the cellular immune response activity, mainly in the initial phase of the infectious process, for posterior involvement of the humoral response, and that both TNF-α and IL-6 could be associated with the immunopathogenesis of the disease.

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%) 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.

Effect of environmental mycobacteria (Mycobacterium avium) on immunity induced by a DNA vaccine (DNAhsp65) against tuberculosis

The efficacy of BCG vaccine (attenuated Mycobacterium bovis) against pulmonary tuberculosis varies enormously among different populations. The prevailing hypothesis attributes this variation to interactions between the vaccine and mycobacteria common in the environment. Studies have revealed that most protective antigens expressed by the antituberculous vaccine are conserved in M. avium, supporting the hypothesis that exposure to environmental mycobacteria generates a cross-reactive immune response that interferes with BCG efficacy. In this study we investigated the effect of a prior exposure to heat-killed M. avium on the immune response and the protective efficacy induced by a genetic vaccine pVAXhsp65 (hsp65 gene from M. leprae inserted in pVAX vector) against experimental tuberculosis. To evaluate the effect on the immune response, female BALB/c mice were initially injected with distinct doses (0.08×106, 4×106, and 200×10 6) of heat-killed M. avium by subcutaneous route. Three weeks later, the animals were immunized with 3 doses of DNAhsp65 by intramuscular route (100μg/15 days apart). Control groups received only M. avium, vaccine (pVAXhsp65), vector (pVAX) or saline solution. Cytokine production and antibody levels were determined by ELISA. To evaluate the effect on the protective efficacy, animals were initially sensitized with 200×106 heat-killed CFU of M. avium by subcutaneous route and then immunized with 3 doses of pVAXhsp65 (100μg/15 days apart) by intramuscular route. Control groups were injected with saline, pVAX (4 doses), pVAXhsp65 (4 doses), M. avium or M. avium plus pVAX (3 doses). Fifteen days after last DNA dose, the animals were infected with 1×104 viable CFU of H37Rv M. tuberculosis by intratracheal route. Thirty days after challenge, the animals were sacrificed and the bacterial burden was determined by counting the number of CFU in the lungs. Lung histological sections were also analyzed. Splenic cells from primed animals produced more IL-5 but less IFN-gamma than non-primed ones. Also, prior contact with M. avium determined higher production of IgG1 and IgG2a anti-hsp65 antibodies in comparison to control groups. However, this higher immune response did not decrease the bacterial burden in the lungs. In addition, prior sensitization with M. avium decreased the parenchyma preservation observed in the group immunized only with pVaxhsp65. These results indicate that environmental mycobacteria can interfere with immunity and protective efficacy induced by DNAhsp65.

Tele-education environment and didactic iconography in sexual transmitted diseases

The field of advertising has developed techniques of mass communication and rapid transmission of information. Among these techniques are slogans and iconographies. This work used such tools for educational purposes and was divided into three modules. Module 1 evaluated the use of simplified textual communication (slogans) about the subject Sexually Transmitted Diseases. It was applied to third-year medical students, which had not taken the course on this theme. To evaluate the impact of the textual communication form (slogans), long phrases were elaborated in scientific language containing 15 topics. From these long phrases, compact phrases were developed using techniques of the advertising area for elaboration of slogans. Three forms of didactic material about that theme were developed. The first form consisted of long phrases in descriptive topics, using scientific language. The second was constituted exclusively of compact phrases in the form of slogans, and the third was composed of the combination of the two previous forms. Then, 10 multiple-choice questions were elaborated and applied in two phases. In the first phase, application occurred immediately after the reading of didactic texts; in the second phase, it happened 60 days after the reading. For statistical analysis, Snedcor's F test was used for analysis of variance, at 5% significance level. There was an increase in memorization by students who read the material containing the association between long phrases and slogans, which indicates that the latter, when used as an auxiliary model of learning, can bring significant benefits for education. Module 2 consisted of analyzing the elaboration of educational videos produced in graphic computing (called iconographies) for development of dynamic communication means. The theme Hair Cycle was utilized. Viability of high quantities of information in few minutes of animation could be demonstrated together with the advantage of presenting the process in a dynamic form without wasting scientific details. Module 3 presented the inclusion of slogans in educational videos produced in graphic computing about the Hair Cycle, showing a new tool for rapid and efficient transference of data. Slogans and iconographies, when utilized in educational material, can bring significant benefits for the student's learning.

Frequency of rhabdiasid nematodes in wild Crotalus durissus terrificus (Serpentes, Viperidae) from Botucatu region, São Paulo State, Brazil

The aim of the present study was to evaluate the frequency of rhabdiasid nematodes in recently captured Crotalus durissus terrificus snakes from São Paulo State, Brazil. Fifty snakes (34 males and 16 females) were studied and each one was evaluated for the presence of that nematode at the moment of receipt at the Institution and after 90 days of quarantine inside individual cages. Tracheopulmonary washes were examined. Snakes that died during quarantine underwent necropsy and lung examination. Analysis of the results obtained at the two evaluation times (0 and 90 days), in addition to the data obtained during necropsies, showed that 44% (18 males and 4 females) of the C. d. terrificus snakes were naturally infected by rhabdiasid nematodes. These data demonstrate the parasitism level in natural conditions and are important for the sanitary handling of these reptiles in captivity.

Intestinal microbiota and HIV-1 infection

The intestinal microbiota consists of a qualitatively and quantitatively diverse range of microorganisms dynamically interacting with the host. It is remarkably stable with regard to the presence of microorganisms and their roles which, however, can be altered due to pathological conditions, diet composition, gastrointestinal disturbances and/or drug ingestion. The present review aimed at contributing to the discussion about changes in the intestinal microbiota due to HIV-1 infection, focusing on the triad infection-microbiota-nutrition as factors that promote intestinal bacterial imbalance. Intestinal microbiota alterations can be due to the HIV-1 infection as a primary factor or the pharmacotherapy employed, or they can be one of the consequences of the disease.

Environmental parameters and antimicrobial susceptibility of enterobacteriaceae isolated from estuarine waters of São Vicente, São Paulo State, Brazil

Diseases transmitted by water consists a serious public health problem and enterobacteria are the main group of microorganisms responsible for outbreaks in humans. Such pathogenic bacteria proliferate in water polluted by domestic and industrial sewage and reach the population through seawater contact. The aim of the present work was to study environmental parameters as well as to identify Enterobacteriaceae species and their antimicrobial susceptibility in water samples collected from the estuarine area of São Vicente city (São Paulo State, Brazil). Strains were identified by using traditional biochemical tests described in literature and antimicrobial susceptibility tests were carried out using the disk diffusion method. Out of 26 samples, Escherichia coli was the most frequent species (40.1%), followed by Citrobacter, Enterobacter and Klebsiella. The most effective drugs against the tested microorganisms were gentamycin, netilmicin, ciprofloxacin and cefepime. Since these bacteria are commonly found in seashore contaminated by sewage effluents, it can be concluded that estuarine waters of São Vicente are polluted and potentially capable of causing diseases and spreading pathogenic bacteria to human communities.

Cytokine profile and clinical metabolic alterations in HIV-1 infected individuals with and without lipodistrophy

The extensive use of Highly Active Antiretroviral Therapy (HAART) has transformed HIV infection into a chronic condition. Thus, metabolic alterations including lipodystrophy and dyslipidemia have been associated with the use of such medications. The objective of the present study was to analyze clinical metabolic alterations and the profile of TNF-α, IFN-γ, IL-2, IL-10, and TNF-α type II soluble receptor in serum of HIV-1 individuals with and without lipodystrophy. Eighty-four adults were evaluated, 42 males and 42 females, mean age 37 years, and HAART time of at least 15 months. Two groups were formed, G1: 42 individuals with lipodystrophy, and G2: 42 without lipodistropy. From the HAART used, stavudine was more associated with the lipodystrophy group and zidovudine with the non-lipodystrophy group. CD4 and CD8 values, viral load, glucose, albumin, and lipids were not different between groups, except for triglycerides, which were high in the lipodystrophy group, and HDL, whose concentration was reduced in G1. TNF-α, TNF-RII, and IL-10 profiles were high and had positive correlation; IL-2 and IFN-γ had reduced levels in the lipodystrophy group. High TNF-α and its receptor levels seem to be associated with lipodystrophy development in individuals under HAART therapy.