Seroprevalence of SARS-CoV-2 in hospital workers in the southern region of Minas Gerais state in Brazil: An analysis of the pre-vaccine period.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection seroprevalence can be performed by detecting anti-SARS-CoV-2 antibodies. The survey is essential to understand the disease transmission's dynamic in the studied population. This study aimed to carry out a seroepidemiological survey of SARS-CoV-2 in three hospitals located in the south of Minas Gerais state, Brazil. 859 samples were collected from August to December 2020 when SARS-CoV-2 vaccines were still not available and Enzyme-linked immunosorbent assays (ELISA) were performed on participants sera. The average age of participants was 38 years, and most were women (71.4%). Likewise, most participants were classified as health professionals with direct or indirect contact with patients with COVID-19 (74.5%). The other participants tested belonged to other sectors, such as the administrative one (11,6%). Considering clinical symptoms, 15.8% of participants reported diarrhoea, 6.4% fever, 5.8% respiratory distress, and 7.0% loss of smell and taste. Many participants reported contact with infected patients (63.35%). Regarding the ELISA tests, 21.6% of the participants had positive results and hospital 3 had the highest positivity (21.7%), followed by hospital 2 (21.6%) and hospital 1 (20.3%). The prevalence was higher in women compared to men (22,8% and 18,7%, respectively). Regarding the area of expertise, the highest positivity (20.9%) was observed among health professionals. However, professionals who worked exclusively with COVID-19 had lower positivity when compared to professionals who did not work directly with COVID-19 (22.0% and 21.5%, respectively). When analysing the correlation between the ELISA tests with the other variables, a significant association was detected with these previous serological variables, previous contact with COVID-19 and the presence of fever symptoms, loss of smell and taste. Clinical symptoms associated with serological tests are important tools for monitoring the disease among health professionals.

Itraconazole and neutrophil interactions in the immune-inflammatory response of paracoccidioidomycosis using a murine air pouch infection model.

Paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis (Pb), is a severe mycosis, prevalent in tropical countries. The presence of polymorphonuclear neutrophils (PMN) in lesions is conspicuous, indicating their central role in innate immunity through the direct killing of Pb and the production of cytokines that activate acquired immunity in the presence of itraconazole (Itra). The toxicity and direct antifungal activity of Itra on Pb in splenocyte co-cultures were evaluated in vitro. Itra showed no toxic effect but marked antifungal activity against Pb. Purified PMN were obtained by the subcutaneous (SC) injection of Pb into mice. Results showed the effect of Itra on the size of the air pouch produced, the cellular population that migrated to the infection site, protein, and mitochondrial metabolism patterns, production of ROS an NO, and the number of cytokines synthesized. Lower doses (3 and 10 mg/kg) of Itra did not affect the cellular profile but led to a lower influx of viable more active PMN, and increased production of ROS and proteins. At a dose of 50 mg/kg the PMN profile remained unchanged along with all other parameters analyzed remained unaltered. Decreases in most cytokine levels were inversely proportional to the Itra concentration. Lower Itra concentrations may elicit activation of the immune response because the combined effects of therapy and immune response are needed, while the higher dose does not require it. Itra also promotes the activation of the cytokines which elicit PMN activation and consequently the resolution of Pb18 infection in the air pouch.

Integrative transcriptome analysis of SARS-CoV-2 human-infected cells combined with deep learning algorithms identifies two potential cellular targets for the treatment of coronavirus disease.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) quickly spread worldwide, leading coronavirus disease 2019 (COVID-19) to hit pandemic level less than 4 months after the first official cases. Hence, the search for drugs and vaccines that could prevent or treat infections by SARS-CoV-2 began, intending to reduce a possible collapse of health systems. After 2 years, efforts to find therapies to treat COVID-19 continue. However, there is still much to be understood about the virus' pathology. Tools such as transcriptomics have been used to understand the impact of SARS-CoV-2 on different cells isolated from various tissues, leaving datasets in the databases that integrate genes and differentially expressed pathways during SARS-CoV-2 infection. After retrieving transcriptome datasets from different human cells infected with SARS-CoV-2 available in the database, we performed an integrative analysis associated with deep learning algorithms to determine differentially expressed targets mainly after infection. The targets found represented a fructose transporter (GLUT5) and a component of proteasome 26s. These targets were then molecularly modeled, followed by molecular docking that identified potential inhibitors for both structures. Once the inhibition of structures that have the expression increased by the virus can represent a strategy for reducing the viral replication by selecting infected cells, associating these bioinformatics tools, therefore, can be helpful in the screening of molecules being tested for new uses, saving financial resources, time, and making a personalized screening for each infectious disease.

Integrative transcriptome analysis of human cells treated with silver nanoparticles reveals a distinct cellular response and the importance of inorganic elements detoxification pathways.

The field of Nanotechnology has taken a great leap in recent decades, with several products currently researched in the industrial sector and even available in the market bringing nanostructured components. The pharmaceutical industry has explored this type of structure as targeted drug delivery, especially against cancer. Integrative transcriptome analysis (ITA) is considered a promising technique for understanding biological events by analyzing several transcriptomes deposited in public databases. This research recovered seven transcriptomes' studies of human cells treated with silver nanoparticles without association or conjugation with any other substance or material for the performance of ITA. This analysis consists of a bipartite network for determining shared differentially expressed genes (DEGs) between different datasets from human cells treated with silver nanoparticles (AgNPs) at both early (4 or 6 h) and late treatment time (24 h). Most of the few upregulated DEGs shared by five or more datasets belong to biological pathways related to mineral absorption, suggesting that these processes were upregulated in AgNPs-treated cells. In addition, Ferroptosis, protein processing in the endoplasmic reticulum, and mitogen-activated protein kinase (MAPK) signaling pathway were also upregulated. Thus, the ITA of human cells treated with AgNPs indicates that the expression profile induced by these nanoparticles is specific to each cell type. However, they share inorganic compounds and oxidative stress responses genes, triggering apoptosis. This work reinforces the need for the biological characterization of cellular response to silver nanoparticles for application in humans, thus ensuring the safety and optimization of the desired results.

Screening paracoccidioidomycosis by double immunodiffusion test in a referred diagnostic center in Brazilian southeastern: an accessible tool.

PURPOSE: This article shows reports of the clinical-epidemiological characteristics and serological screening in patients assisted by a reference center for PCM care, University Hospital Cassiano Antonio Moraes, Federal University of Espirito Santo, Brazil. METHODS: The patient's sera with PCM were analyzed by DID test at the beginning and the end treatment. Clinical and demographic data were also collected to characterize the sample. RESULTS: One hundred patients with a suspected diagnosis of PCM were evaluated. Serology by DID test was used as a screen in all patients. The test was positive for 79 patients (72 for Paracoccidioides brasiliensis and 7 for Paracoccidioides lutzii). Serology was negative in 21 sera, although all of them were diagnosed PCM by histopathologic or direct exam. Serological follow-up was performed during the treatment of all patients. After treatment, 58(58%) had negative serology and 33(33%) low levels of antibodies (≤ 1:16). CONCLUSION: Our results indicate the importance of the DID test for the screening and monitoring of PCM and that the incidence of P. lutzii might be greater than expected in areas where it is not the predominant PCM species. Therefore, this article may contribute to improving the knowledge and clinical management about this disease.

Nanoparticles as Vaccines to Prevent Arbovirus Infection: A Long Road Ahead.

Arthropod-borne viruses (arboviruses) are a significant public health problem worldwide. Vaccination is considered one of the most effective ways to control arbovirus diseases in the human population. Nanoparticles have been widely explored as new vaccine platforms. Although nanoparticles' potential to act as new vaccines against infectious diseases has been identified, nanotechnology's impact on developing new vaccines to prevent arboviruses is unclear. Thus, we used a comprehensive bibliographic survey to integrate data concerning the use of diverse nanoparticles as vaccines against medically important arboviruses. Our analysis showed that considerable research had been conducted to develop and evaluate nanovaccines against Chikungunya virus, Dengue virus, Zika virus, Japanese encephalitis virus, and West Nile virus. The main findings indicate that nanoparticles have great potential for use as a new vaccine system against arboviruses. Most of the studies showed an increase in neutralizing antibody production after mouse immunization. Nevertheless, even with significant advances in this field, further efforts are necessary to address the nanoparticles' potential to act as a vaccine against these arboviruses. To promote advances in the field, we proposed a roadmap to help researchers better characterize and evaluate nanovaccines against medically important arboviruses.

Seroepidemiological survey on sporotrichosis-infection in rural areas of the south of Minas Gerais State, Brazil.

Sporotrichosis is a subcutaneous mycosis caused by traumatic inoculation into the skin by fungi species of the genus Sporothrix. The disease has different clinical manifestations (cutaneous, lymphocutaneous, and disseminated), and can also progress to a systemic infection. Despite having a worldwide distribution, sporotrichosis is most prevalent in tropical and subtropical countries. In Brazil, reports of the disease are higher frequent, where cases of the disease were found in Rio de Janeiro, Sao Paulo, Curitiba, Pernambuco, and Paraiba, among others. Certain groups of people may be more exposed to the causative agent of disease, such as residents of rural areas. Thus, this work aimed to carry out a seroepidemiological survey of the prevalence of sporotrichosis in four rural locations in the south of Minas Gerais State, Brazil. In this study, we used an indirect ELISA test in the survey on the prevalence of sporotrichosis. Data obtained in this study evaluated a population of 631 individuals and showed a prevalence of 44.69%. The distribution of seroprevalence of sporotrichosis with respect to age groups and gender showed no significant statistical difference. Thus, we found a high seroprevalence of sporotrichosis-infection in rural regions of southern Minas Gerais State, Brazil, with no difference in prevalence in relation to gender and age.

Detection of Serum Antibodies and Peripheral Blood Mononuclear Cells Response to Escherichia coli Antigens in Humans

Abstract Among the microorganisms that make up the intestinal microbiota, stands out Escherichia coli, which has as main ecological niche, the large human intestine. Its importance stands out in being part of the pioneer's commensal microorganisms on the colonization of the intestinal mucosa and its pathogenic role causing extra and intra intestinal diseases. The aim of the study was to evaluate the antibody production and proliferative response of Peripheral Blood Mononuclear Cells (PBMC) to E. coli antigens. The bacteria were grown on Brain Heart Infusion broth medium at 35 ºC for 72 hours. Pellet bacteria were lysed for one hour at room temperature with an 8M sodium guanidine solution. After spin and dialysis, the protein antigens were measured in the supernatant by protein assay. The antigens were characterized by polyacrylamide gel electrophoresis and the antigenic profile by western blotting. The presence of specific IgG and IgA antibodies were evaluated using thirty normal human sera by an indirect ELISA. The response of PBMC to E. coli antigens was assessed by MTT metabolization. The results demonstrated that the antigens were composed of proteins of different sizes and they were recognized by antibodies present in normal human serum. Human sera presented high titers of IgG and IgA antibodies to E. coli antigens when compared to the results of lipopolysaccharide. We also showed that total E. coli antigens induced PBMC proliferation at different antigen concentrations. Taken together the results suggest that the antigens from E. coli can induce local and systemic immune responses.

Medication association and immunomodulation: An approach in fungal diseases and in particular in the treatment of paracoccidioidomycosis.

Fungal infections have been increasing in recent decades, mainly affecting immunocompromised individuals, although certain mycoses, such as paracoccidioidomycosis (PCM), infect immunologically competent individuals. The major problems observed regarding fungal diseases are inadequate diagnosis, prolonged treatment time, the reduced number of drugs available for treatment, in addition to the fact that there are no vaccines for clinical use. Drug combination in order to immunomodulate the immune response is a new strategy used for the treatment of mycoses, since it is difficult to develop new antifungal drugs. The aim of this study is to present and analyze strategies recently suggested for the treatment of fungi of medical interest, in particular for PCM, such as the utilization of combinations of protein fractions or dead microorganisms, as vaccinal antigens, and cellular immunotherapy. We will also propose new therapeutic alternatives, such as lipids, vitamins, synthetic or natural products as well as the use of low intensity LASER therapy (LLLT) to modulate the immune response of the host, enhancing the efficiency of the existing treatments of mycoses of medical interest and in particular of PCM.

Virulence factors of Paracoccidioides brasiliensis as therapeutic targets: a review.

Paracoccidiodomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis and Paracoccidioides lutzii. The disease requires long and complicated treatment. The aim of this review is to address the fungal virulence factors that could be the target of the development of new drugs for PCM treatment. Virulence factors favoring the process of fungal infection and pathogenicity are considered as a microbial attribute associated with host susceptibility. P. brasiliensis has some known virulence factors which are 43 kDa glycoprotein (gp 43) which is an important fungal antigen, 70 kDa glycoprotein (gp 70), the carbohydrates constituting the fungal cell wall α-1,3, glucan and ß-1,3-glucan, cell adhesion molecules and the presence of melanin pigments. The discovery and development of drugs that interact with these factors, such as inhibitors of ß-1,3-glucan, reduced synthesis of gp 43, inhibitors of melanin production, is of great importance for the treatment of PCM. The study of virulence factors favors the understanding of pathogen-host relationships, aiming to evaluate the possibility of developing new therapeutic targets and mechanisms that these molecules play in the infectious process, favoring the design of a more specific treatment for this disease.

Correction to: Effects of metaperiodate and urea solutions on the serological diagnosis of human sporotrichosis using an indirect ELISA test.

In the article mentioned above an author's name was misspelled. The correct author name reads as follows: Leila Maria Lopes-Bezerra. We apologize for the inconvenience.

Effects of metaperiodate and urea solutions on the serological diagnosis of human sporotrichosis using an indirect ELISA test.

Sporotrichosis is an infection of the skin caused by traumatic inoculation of the fungus Sporothrix schenckii. Definitive diagnosis relies on direct visualization of the fungus or its isolation on culture medium, although both have low sensitivity. Alternatively, the detection of the antibody response offers a more rapid alternative for diagnosis. Although the available immunoassays possess good sensitivity and specificity, cross-reactivity is still a problem. This study aimed to evaluate the effect of sodium metaperiodate and 6 M urea solutions on the serological diagnosis of sporotrichosis using an enzyme-linked immunosorbent assay (ELISA) test. Ninety-six-well plates were sensitized with exoantigens from the yeast phase of S. schenckii. Sera of patients with confirmed sporotrichosis, sera of patients with paracoccidioidomycosis, and sera of individuals with a sporotrichin-negative skin test were tested. Two strategies were used; the first consisted of treating the antigen with sodium metaperiodate solution for different incubation times, and the second consisted of treating the serum with 6 M urea solution for different incubation times. ROC curve analysis revealed that the best discrimination parameters were obtained using 6 M urea solution incubated for 5 min and serum dilution at 1/600. The use of 6 M urea solution improves the performance of the ELISA test in the diagnosis of sporotrichosis.

DC-SIGN and VDR polymorphisms are associated with chronic form of paracoccidioidomycosis with oral manifestations.

Paracoccidioidomycosis (PCM) is a granulomatous disease caused by fungi of the species complex of the Paracoccidioides genus. One of the main clinical manifestations of PCM is the presence of oral lesions with the presence of epithelioid granulomas. In this work, we aimed to evaluate the frequency of SNPs in the TNF-α, JAK1, VDR, DC-SIGN and FcγRIIa genes in patients with chronic PCM and verify possible association of these SNPs with the organisation pattern of the granulomas in the oral lesions. A total of 66 samples of DNA were obtained from oral lesions biopsies and 106 DNA samples were obtained from healthy individuals. The individuals were genotyped for SNPs in DC-SIGN (rs4804803), FcγRIIa (rs1801274), JAK1 (rs11208534), TNF-α (rs1800629) and VDR (rs7975232) by real-time PCR and allele discrimination method. Granulomas were classified as loose or dense according to the histological pattern. In the VDR (rs7975232), the CC genotype (P < 0.001, OR = 5.94, 95% CI = 2.07-17.05), and the C allele (P = 0.027, OR = 2.71, 95% CI = 1.07-6.86), as well as the GG genotype in DC-SIGN (rs4804803) (P = 0.032, OR: 3.76, 95%, I = 1.06-13.38) are associated with an increased risk of oral PCM. Our data indicate that VDR and DC-SIGN genetics variations are related to the susceptibility of oral PCM in the group of patients analysed.

Bovine serum albumin nanoparticles induce histopathological changes and inflammatory cell recruitment in the skin of treated mice.

Albumin is a natural, biocompatible, biodegradable and nontoxic polymer and due to these features, nanoparticles made of albumin are a good system for drug or antigen delivery. Polymeric nanoparticles are being widely explored as new vaccines platforms due to the capacity of those nanoparticles to prime the immune system by providing sustained release of the antigen after injection. Biodegradable nanoparticles associated with proteins represent a promising method for in vivo delivery of vaccines. In our previous studies, bovine serum albumin nanoparticles (BSA-NPs) were identified as a promising system for in vivo delivery of microbial antigens. The aim of this work was to show the effect of BSA-NPs on skin after nanoparticles administration. The pro-inflammatory activity of BSA-NPs was evaluated using in vivo models. BSA-NPs are easily uptake by macrophagic RAW 264.7 and BHK-21 cells without any significant cytotoxicity. Histological examination of skin sections from BSA-NPs-treated mice revealed intense cellular infiltration, increased skin thickness, follicular hypertrophy, vascular congestion and marked collagenesis. Mice immunized with recombinant non-structural protein 1 (rNS1) from Dengue virus 1 and BSA-NPs showed a high seroconversion rate if compared to animals immunized only with rNS1. Therefore, the effect of BSA-NPs on skin after BSA-NPs administration has a biotechnological relevance to the rational design of vaccine formulations based on albumin nanocarriers. However in the next years future studies should be carried out to best characterize the effect of BSA-NPs on dendritic cells and establish the role of these nanoparticles as a new vaccine platform for infectious diseases or cancer.

High prevalence of dengue antibodies and the arginine variant of the FcγRIIa polymorphism in asymptomatic individuals in a population of Minas Gerais State, Southeast Brazil.

Dengue is the most prevalent arthropod-borne viral illness in humans worldwide. Single-nucleotide polymorphisms (SNPs) in genes involved in the immune response, such as dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN), IgG Fc receptor II-A (FcγRIIa), vitamin D receptor (VDR), and tumor necrosis factor alpha (TNF-α), were previously reported to be associated with susceptibility to dengue disease in different human populations. Therefore, due to the relevant association of host immune and genetic status with disease susceptibility/severity of dengue, this work aims to verify the frequency of anti-dengue virus antibodies and some dengue-associated risk SNPs in a population in Minas Gerais State, Southeast Brazil. A total of 1560 individuals were genotyped for polymorphisms in DC-SIGN (rs4804803), FcγRIIa (rs1801274), VDR (rs7975232), and TNF-α (rs1800629). The presence of anti-dengue antibodies (IgM and/or IgG) in these samples was also assayed. Anti-dengue antibodies were detected at an overall frequency of 16.86%, indicating a virus infection in asymptomatic individuals. The genotypic frequencies of all SNPs studied did not differ between the asymptomatic and control groups. Regarding the allelic frequencies of the four SNPs analyzed, a higher frequency was detected of the G allele of FcγRIIa/rs1801274 in the asymptomatic individuals when compared to that in the control group (p = 0.03). Therefore, the results showed a high prevalence of asymptomatic individuals in Minas Gerais State, with a potential association between the presence of the G allele of FcγRIIa/rs1801274 and protection against symptomatic disease.

Phylogenetic analysis of Dengue virus 1 isolated from South Minas Gerais, Brazil

Abstract Dengue is a major worldwide public health problem, especially in the tropical and subtropical regions of the world. Primary infection with a single Dengue virus serotype causes a mild, self-limiting febrile illness called dengue fever. However, a subset of patients who experience secondary infection with a different serotype can progress to a more severe form of the disease, called dengue hemorrhagic fever. The four Dengue virus serotypes (1–4) are antigenically and genetically distinct and each serotype is composed of multiple genotypes. In this study we isolated one Dengue virus 1 serotype, named BR/Alfenas/2012, from a patient with dengue hemorrhagic fever in Alfenas, South Minas Gerais, Brazil and molecular identification was performed based on the analysis of NS5 gene. Swiss mice were infected with this isolate to verify its potential to induce histopathological alterations characteristic of dengue. Liver histopathological analysis of infected animals showed the presence of inflammatory infiltrates, hepatic steatosis, as well as edema, hemorrhage and necrosis focal points. Phylogenetic and evolutionary analyses based on the envelope gene provided evidence that the isolate BR/Alfenas/2012 belongs to genotype V, lineage I and it is probably derived from isolates of Rio de Janeiro, Brazil. The isolate BR/Alfenas/2012 showed two unique amino acids substitutions (SER222THRE and PHE306SER) when compared to other Brazilian isolates from the same genotype/lineage. Molecular models were generated for the envelope protein indicating that the amino acid alteration PHE 306 SER could contribute to a different folding in this region located within the domain III. Further genetic and animal model studies using BR/Alfenas/2012 and other isolates belonging to the same lineage/genotype could help determine the relation of these genetic alterations and dengue hemorrhagic fever in a susceptible population.

Phylogenetic analysis of Dengue virus 1 isolated from South Minas Gerais, Brazil.

Dengue is a major worldwide public health problem, especially in the tropical and subtropical regions of the world. Primary infection with a single Dengue virus serotype causes a mild, self-limiting febrile illness called dengue fever. However, a subset of patients who experience secondary infection with a different serotype can progress to a more severe form of the disease, called dengue hemorrhagic fever. The four Dengue virus serotypes (1-4) are antigenically and genetically distinct and each serotype is composed of multiple genotypes. In this study we isolated one Dengue virus 1 serotype, named BR/Alfenas/2012, from a patient with dengue hemorrhagic fever in Alfenas, South Minas Gerais, Brazil and molecular identification was performed based on the analysis of NS5 gene. Swiss mice were infected with this isolate to verify its potential to induce histopathological alterations characteristic of dengue. Liver histopathological analysis of infected animals showed the presence of inflammatory infiltrates, hepatic steatosis, as well as edema, hemorrhage and necrosis focal points. Phylogenetic and evolutionary analyses based on the envelope gene provided evidence that the isolate BR/Alfenas/2012 belongs to genotype V, lineage I and it is probably derived from isolates of Rio de Janeiro, Brazil. The isolate BR/Alfenas/2012 showed two unique amino acids substitutions (SER222THRE and PHE306SER) when compared to other Brazilian isolates from the same genotype/lineage. Molecular models were generated for the envelope protein indicating that the amino acid alteration PHE 306 SER could contribute to a different folding in this region located within the domain III. Further genetic and animal model studies using BR/Alfenas/2012 and other isolates belonging to the same lineage/genotype could help determine the relation of these genetic alterations and dengue hemorrhagic fever in a susceptible population.

Demethylation profile of the TNF-α promoter gene is associated with high expression of this cytokine in Dengue virus patients.

Dengue is the most prevalent arthropod-borne viral illness in humans. The overexpression of cytokines by Dengue virus (DENV) infected cells is associated with the most severe forms of the disease. Unmethylated CpG islands are related to a transcriptionally active structure, whereas methylated DNA recruits methyl-binding proteins that inhibit gene expression. Several studies have described the importance of epigenetic events in the regulation and expression of many cytokines. The purpose of the present study was to evaluate the methylation status of the IFN-γ and TNF-α promoters in DNA extracted from dengue infected patients using methylation-specific polymerase chain reaction. A high frequency of demethylation was observed in the TNF-α promoter of DENV infected patients when compared to non-infected controls. The patients with an unmethylated profile showed higher expression of TNF-α mRNA than patients with the methylated status. No difference was found in the methylation frequency between the two analyzed groups regarding the IFN-γ promoter or in the expression of IFN-γ transcripts. The present study provides the first association of TNF-α promoter demethylation in DENV infected individuals and demonstrates a correlation between the methylation status of the region analyzed and the expression of TNF-α transcripts in DENV infected patients. J. Med. Virol. 88:1297-1302, 2016. © 2016 Wiley Periodicals, Inc.

Serum antibody levels correlate with oral fungal cell numbers and influence the patients' response to chronic paracoccidioidomycosis.

Paracoccidioidomycosis (PCM) is a neglected fungal disease that elicits an important granulomatous inflammatory reaction which aims to isolate the fungi and resolve the infection; besides the innate cellular response, the patients' sera may contain different levels of antibodies directed against PCM's pathogenic agent: Paracoccidioides brasiliensis (Pb). The aim of the study was to assess the distinct serum antibody levels of 19 chronic PCM patients and to associate these levels to the granulomatous inflammatory response and presence of fungi in oral lesions caused by Pb. The presence of Pb was detected and counted within oral tissues using immunohistochemistry; antibody levels were classified as negative, low-grade, moderate or high-grade groups. The Kruskal-Wallis and Dunn's test were used to verify possible associations among the groups. Interestingly, lower antibody titres were associated with lesser numbers of Pb, which favours the cellular response over the humoral response to fight PCM. On the other hand, negative serological results were linked to a higher presence of Pb in the tissues, indicating that a deficient humoral response supports the fungal proliferation. The number of Pb was conveniently associated with the level of serum antibodies, showing that the humoral immune response is required, however, not solely responsible to restrain the dissemination of Pb.

Prevalence of paracoccidioidomycosis infection by intradermal reaction in rural areas in Alfenas, Minas Gerais, Brazil.

This study aimed to estimate the prevalence of paracoccidioidal infection by intradermal reaction (Delayed-Type Hypersensitivity, DTH) to Paracoccidioides brasiliensis in rural areas in Alfenas, Southern Minas Gerais (MG) State, Brazil, and to assess risk factors (gender, occupation, age, alcohol intake and smoking) associated with infection. We conducted a population-based cross-sectional study using intradermal tests with gp 43 paracoccidioidin in 542 participants, who were previously contacted by local health agents and so spontaneously attended the test. Participants underwent an interview by filling out a registration form with epidemiological data and were tested with an intradermal administration of 0.1 mL of paracoccidioidin in the left forearm. The test was read 48 hours after injection and was considered positive if induration was greater than or equal to 5 mm. Out of 542 participants, 46.67% were positive to the skin test. Prevalence increased in accordance with an increase of age. There was statistical significance only for males. Occupation, alcohol intake and smoking habits were not significantly associated with the risk of paracoccidioidomycosis infection. There is relevance of paracoccidioidomycosis infection in such rural areas, which suggests that further epidemiological and clinical studies on this mycosis should be done in the southern part of Minas Gerais State.