Diagnosis of hepatitis C virus in Brazilian blood donors using a reverse transcriptase nested polymerase chain reaction: comparison with enzyme immunoassay and recombinant protein immunoblot assay

Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA > or = 3, ALT<1), group B (EIA > or = 3, ALT>1), group C (1<=EIA<3, ALT<1), group D (1<=EIA<3, ALT>1) and group E (EIA<=0.7). HCV sequences were detected by RT-nested PCR, using primers for the most conserved region of viral genome. RIBA-2 was applied to the same samples. In group A (n=6), all samples were positive by RT-nested PCR and RIBA-2. Among 124 samples in group B, 120 (96.8%) were RIBA-2 positive and 4 (3.2%) were RIBA-2 indeterminate but were seropositive for antigen c22.3. In group B, 109 (87.9%) of the RIBA-2 positive samples were also RT-nested PCR positive, as well as were all RIBA-2 indeterminate samples. In group C, all samples (n=9) were RT-nested PCR negative: 4 (44.4%) were also RIBA-2 negative, 4 (44.4%) were RIBA-2 positive and 1 (11.1%) was RIBA-2 indeterminate. HCV-RNA was detected by RT-nested PCR in 3 (37.5%) out of 8 samples in group D. Only one of them was also RIBA-2 positive, all the others were RIBA-2 indeterminate. All of the group E samples (controls) were RT- nested PCR and RIBA-2 negative. Our study suggests a strong relation between anti-HCV EIA-2 ratio > or = 3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR (75%), especially when the c22.3 band was detected.

Hantavirus pulmonary syndrome in Anajatuba, Maranhão, Brazil

The authors report a confirmed case of hantavirus pulmonary syndrome in the rural area of the municipality of Anajatuba, state of Maranhão. Two other suspected cases from the same region are also described. The confirmed case involved a previously healthy young woman who died with signs and symptoms of acute respiratory insufficiency 5 days after presenting fever, myalgia and a dry cough. The patient was a student who was helping her parents with work in the fields; it was a habit of the family to store rice inside the house. The suspected cases involved two first-degree relatives working as field hands who died of acute respiratory insufficiency 24 and 48 hours, respectively, after presenting fever, myalgia and a dry cough. Both stored rice and corn inside their home. People living in the region reported massive infestations with rats in the woods and fields.

Liver histology in co-infection of hepatitis C virus (HCV) and Hepatitis G virus (HGV)

As little is known about liver histology in the co-infection of hepatitis C virus (HCV) and hepatitis G virus (HGV), HGV RNA was investigated in 46 blood donors with hepatitis C, 22 of them with liver biopsy: co-infection HCV / HGV (n = 6) and HCV isolated infection (n = 16). Besides staging and grading of inflammation at portal, peri-portal and lobular areas (Brazilian Consensus), the fibrosis progression index was also calculated. All patients had no symptoms or signs of liver disease and prevalence of HGV / HCV co-infection was 15.2%. Most patients had mild liver disease and fibrosis progression index, calculated only in patients with known duration of infection, was 0.110 for co-infection and 0.130 for isolated HCV infection, characterizing these patients as "slow fibrosers". No statistical differences could be found between the groups, although a lesser degree of inflammation was always present in co-infection. In conclusion co-infection HCV / HGV does not induce a more aggressive liver disease, supporting the hypothesis that HGV is not pathogenic.

Intestinal parasitoses and environmental factors in a rural population of Argentina, 2002-2003

We evaluate the prevalence of intestinal parasites in 504 people and the degree of association between environmental variables and parasites found in population, soil and water in a rural area of Argentina during 2002-2003. A structured survey was used to evaluate the environmental variables and fecal-human, soil and water samples were analyzed. The prevalence of parasites was 45.4%. Most prevalent protozoa were Blastocystis hominis (27.2%) and Giardia lamblia (6.9%), while the most prevalent helminth was Ascaris lumbricoides (3.8%). The analyzed environmental variables showing association (p < 0.05) with presence of parasites in population were: cardboard-tin or wooden house, dirt floor, home or communal water pump, faucet outside the house or public faucet and cesspool or latrine. Parasite forms were found in 82.3% of the soil samples and in 84.2% of the water samples. In both samples we found parasites that were also found in people. In this study we have found deficient sanitary conditions associated with presence of parasites in population and we have evidenced that contaminated soil and water were the source of these parasites.

Assessment of physical impairments in leprosy patients: a comparison between the world health organization (who) disability grade and the Eye-Hand-Foot Score

This cross-sectional study assessed the grade of physical impairments in 61 individuals with leprosy receiving multidrug therapy (MDT) under the Brazilian Unified Health System (SUS), and residing in Campina Grande, Paraíba State, Brazil. Impairments were assessed using the disability grade (DG) standardized by the WHO, and the EHF score (Eye-Hand-Foot sum of impairment scores). Impairments were detected in 25 (41%) of the subjects. A total of 14 (23%) patients scored DG 1, while 11 (18%) were assigned DG 2. The EHF score ranged from 1 to 10 points in the group of patients with physical impairments, with a mean score of 3.6 points. The majority of individuals with impairments were affected in at least two sites. We conclude that the EHF score showed overlapping impairments in the segments examined and may be more appropriate than the DG classification system for describing the degree of physical impairment of leprosy patients.

High prevalence of dna from non-H. pylori helicobacters in the gastric mucosa of venezuelan pet dogs and its histological alterations

Non-H. pylori helicobacters (NHPH) have been demonstrated as gastric spiral-shaped bacteria in specimens obtained from dogs; however, their roles in the pathogenesis of upper gastrointestinal disease have not yet been clearly established. The purpose of this study was to evaluate the prevalence of NHPH DNA in the gastric mucosa of dogs and its association with histopathology. Helicobacter was detected through histopathological techniques, PCR, and FISH analysis from fundic biopsies of twenty dogs with or without signs of gastrointestinal disease. PCR and FISH were based on partial 16S rRNA gene sequences. Nineteen dogs showed mild to marked gastritis in the fundus, and only one dog had a healthy gastric mucosa. NHPH DNA was detected in 18 dogs with gastritis and one with normal gastric mucosa. However, there was no significant correlation between the presence of NHPH DNA and the degree of gastritis. These results show a high prevalence of NHPH DNA in the gastric mucosa of dogs from Venezuela. Further studies are necessary to determine a possible association between a specific NHPH species and the degree of gastritis.

Environmental analyses of the parasitic profile found in the sandy soil from the Santos municipality beaches, SP, Brazil

The environmental contamination by geohelminths represents a world public health problem and has been well documented by several authors. However, few papers describe the presence of such contamination in saline soils of coastal beaches. A study was performed on the beaches of the municipality of Santos in the period between May 2004 to April 2005 with the aim of determining the degree of contamination, and the correlation between contamination level and seasonal conditions and characteristics of the environment. Of the 2,520 samples analyzed, 18.2% (458) were contaminated, 32.3% (148) of which were localized in children's recreational areas (playgrounds). The parasite profile found in the analyzed samples indicated the presence of several zoonotic parasites: Ancylostoma larvae (82.5%), Toxocara sp. eggs (59.4%), Ancylostomidae-like eggs (37.1%), coccid oocysts (13.5%), Trichostrongylus sp. eggs and larvae, Ascaris lumbricoides eggs, (11.6%), Entamoeba sp. cysts (10.0%), Strongyloides sp. (4.8%), several free nematoids and some non-identified parasitic structures (3.3%). It was established that the highest frequency of parasitic structures occurred in the months between May and October 2004, and from February to March 2005. An increase in the diversity of parasitic forms was documented in the months between February to December 2004 and from January to April 2005, these periods having the highest rainfall.

Rhodococcus equi infection after reduction mammaplasty in an immunocompetent patient

The majority of infections caused by R. equi occur in hosts with some degree of cell-mediated immunodeficiency. Immunocompetent individuals are infrequently affected and usually present with localized disease. Infections of the skin or related structures are uncommon and are usually related to environmental contamination. The microbiology laboratory plays a key role in the identification of the organism since it may be mistaken for common skin flora. We describe a 31 year-old woman without medical problems who presented nine weeks after breast reduction with right breast cellulitis and purulent drainage from the surgical wound. She underwent incision and drainage, and cultures of the wound yielded Rhodococcus equi. The patient completed six weeks of antimicrobial therapy with moxifloxacin and rifampin with complete resolution.

Scientific authorships and collaboration network analysis on Chagas disease: papers indexed in PubMed (1940-2009)

Chagas disease is a chronic, tropical, parasitic disease, endemic throughout Latin America. The large-scale migration of populations has increased the geographic distribution of the disease and cases have been observed in many other countries around the world. To strengthen the critical mass of knowledge generated in different countries, it is essential to promote cooperative and translational research initiatives. We analyzed authorship of scientific documents on Chagas disease indexed in the Medline database from 1940 to 2009. Bibliometrics was used to analyze the evolution of collaboration patterns. A Social Network Analysis was carried out to identify the main research groups in the area by applying clustering methods. We then analyzed 13,989 papers produced by 21,350 authors. Collaboration among authors dramatically increased over the study period, reaching an average of 6.2 authors per paper in the last five-year period. Applying a threshold of collaboration of five or more papers signed in co-authorship, we identified 148 consolidated research groups made up of 1,750 authors. The Chagas disease network identified constitutes a "small world," characterized by a high degree of clustering and a notably high number of Brazilian researchers.

Effects of vitamin C supplementation on acute phase Chagas disease in experimentally infected mice with Trypanosoma cruzi QM1 strain

The tissue changes that occur in Chagas disease are related to the degree of oxidative stress and antioxidant capacity of affected tissue. Studies with vitamin C supplementation did not develop oxidative damage caused by Chagas disease in the host, but other studies cite the use of peroxiredoxins ascorbate - dependent on T. cruzi to offer protection against immune reaction. Based on these propositions, thirty "Swiss" mice were infected with T. cruzi QM1 strain and treated with two different vitamin C doses in order to study the parasitemia evolution, histopathological changes and lipid peroxidation biomarkers during the acute phase of Chagas disease. The results showed that the parasite clearance was greater in animals fed with vitamin C overdose. There were no significant differences regarding the biomarkers of lipid peroxidation and inflammatory process or the increase of myocardium in animals treated with the recommended dosage. The largest amount of parasite growth towards the end of the acute phase suggests the benefit of high doses of vitamin C for trypomastigotes. The supplementation doesn't influence the production of free radicals or the number of amastigote nests in the acute phase of Chagas disease.

AN UPWARD TREND IN DNA P16INK4A METHYLATION PATTERN AND HIGH RISK HPV INFECTION ACCORDING TO THE SEVERITY OF THE CERVICAL LESION

SUMMARY High-risk human papillomavirus (hr-HPV) infection is necessary but not sufficient for cervical cancer development. Recently, P16INK4A gene silencing through hypermethylation has been proposed as an important cofactor in cervical carcinogenesis due to its tumor suppressor function. We aimed to investigate P16INK4A methylation status in normal and neoplastic epithelia and evaluate an association with HPV infection and genotype. This cross-sectional study was performed with 141 cervical samples from patients attending Hospital Moncorvo Filho, Rio de Janeiro. HPV detection and genotyping were performed through PCR and P16INK4A methylation by nested-methylation specific PCR (MSP). HPV frequency was 62.4% (88/141). The most common HPV were HPV16 (37%), HPV18 (16.3%) and HPV33/45(15.2%). An upward trend was observed concerning P16INK4A methylation and lesion degree: normal epithelia (10.7%), low grade lesions (22.9%), high grade (57.1%) and carcinoma (93.1%) (p < 0.0001). A multivariate analysis was performed to evaluate an association between methylation, age, tobacco exposure, HPV infection and genotyping. A correlation was found concerning methylation with HPV infection (p < 0.0001), hr-HPV (p = 0.01), HSIL (p < 0.0007) and malignant lesions (p < 0.0001). Since viral infection and epigenetic alterations are related to cervical carcinoma, we suggest that P16INK4A methylation profile maybe thoroughly investigated as a biomarker to identify patients at risk of cancer.

The effect of phenolated "vaccines" against experimental T. cruzi infection in mice

"Vaccines" prepared from parasites of an avirulent cultivated Y strain of T. cruzi, suspended in phenolated 1/10.000 saline solution, with aluminum stearate, containing alive parasites, gave high degree of protection to mice against a posterior infection with virulent blood forms of the same parasites and strain. The degree of protection with 1/1000 and 1/10.000 phenol "vaccines", with no alive parasites, was very poor specially in the first group. The immunity seems to be related to the number of alive trypanosomes in the "vaccines".

Experimental paracoccidioidomycosis in the mouse. III. Histopathological and immunological findings after intravenous infection in the presence or absence of previous immunization

Fifty male white Swiss mice aged 4 weeks were inoculated with 5 x 10(5) viable yeast forms of Paracoccidioides brasiliensis (strain 18). Ten of these animals had been previously immunized with particulate P. brasiliensis antigenfor 4 weeks by intradermal injection. The controls consisted of 10 animals that were only immunized and 10 animals submitted to no treatment. The animals were sacrificed 2, 4, 7,11 and 16 weeks later. We studied: 1) the anti-P. brasiliensis delayed hypersensitivity response measured by the footpad test 24 hours prior to sacrifice; 2) the specific antibody production measured by double immunodiffusion in agar gel; 3) the histopathology of lungs, liver, spleen, adrenals and kidneys. We observed that: a) the immunized animals developed more intense cell-immune responses than the infected ones; b) infection reduced the cell- immune response of the immunized animals; c) intravenous infection of mice with P. brasiliensis was characterized by a systemic and progressive granulomatous inflammation. The animals infected after previous immunization showed less extensive lung inflammation, with smaller granulomas and fewer fungi. The results indicate that the present murine model mimics some findings of the human subacute form of paracoccidioidomycosis (systemic disease with depressed cellular immunity) and that the extrapulmonary immunization scheme was able to induce a certain degree of protection of the lung from infection with P. brasiliensis

Environmental contamination by Toxocara sp eggs in public areas of Salvador, Bahia state, Brazil

A survey ofparasite eggs and cysts in soil and dog feces collected in public places of 23 boroughs of Salvador, a city in the Northeast of Brazil, was performed. High degree of contamination by Toxocara sp eggs was observed in all boroughs studied; other parasites found included: Ascaris lumbricoides, hookworms, whipworms and protozoan cysts. Parks andpublic gardens were more contaminated than streets and beachesfor all parasites, including Toxocara sp.

The hamster (mesocricetus auratus) as experimental model in chagas' disease: parasitological and histopathological studies in acute and chronic phases of Trypanosoma cruzi infection

This research characterizes the acute and chronic phases of Chagas ' disease in hamster through parasitological and histopathological studies. The acute phase was achieved with 44 young hamsters injected intraperitoneally with 100.000 blood trypomastigotes of Benedito and Y strains of T. cruzi. The chronic phase was induced in 46 hamsters injected intraperitoneally with 35.000 trypomastigotes ofVicentina, Benedito and Y strains. Animals were sacrificed at regular intervals of 24 hours of acute phase and from the 3rd to the 10th month of infection ofchronic phase. In the acute phase, parasites were easily recoveredfrom all animals and there was an inflammatory reaction characterized by mononuclear and polymorphous leukocyte infiltration of variable degree in the majority of tissues and organs, specially in the connective loose and fatty tissues, smooth muscle myocardium and skeletal muscle. In the chronic phase the lesions occurred in the same tissues and organs, but the inflammatory response was less severe and characterized by mononuclear infiltration mainly with focal or zonalfibrosis in the myocardiun. In 50% of infected animals parasites were found inmyocardiun and recoveredfrom pericardic, peritoneal and ascitic fluids in some animals. Signs of heart failure, sudden death and enlargement of bowel were observed regularly. We concluded that the hamster is an useful model for Chagas' disease studies.