Clinical comparison of whitening efficacy and tooth sensitivity of different concentrations of hydrogen peroxide photoactivated with violet or blue LEDs.

This study evaluated the photoactivation of hydrogen peroxide gels at different concentrations using blue or violet LED in terms of whitening efficacy and tooth sensitivity. Forty patients were randomly divided into 4 groups: HP6V (violet LED and 6% hydrogen peroxide), HP6B (blue LED and 6% hydrogen peroxide), HP35V (violet LED and 35% hydrogen peroxide), and HP35B (blue LED and 35% hydrogen peroxide). The L*, a* and b* values were measured before, 1 week and 3 months after treatment, and the ΔE and ΔWID values were calculated. Tooth sensitivity was measured using a visual analogue scale (VAS) before, immediately after, and 24 h after bleaching. The ΔE, ΔWID and bleaching sensitivity values were subjected to the ANOVA test and Bonferroni post-test. HP35V and HP35B showed higher whitening efficacy than HP6VL, while HP6V did not show statistical differences compared to the other groups. Regarding bleaching-related sensitivity, the HP6V and HP6B groups presented the lowest values when compared to HP35V and HP35B. HP6V showed whitening efficacy comparable to HP35V and HP35B but with reduced tooth sensitivity. TRIAL REGISTRATION NUMBER: NCT06165458; registration date: 12/09/2023.

Detection of early-stage lung cancer in sputum using automated flow cytometry and machine learning.

BACKGROUND: Low-dose spiral computed tomography (LDCT) may not lead to a clear treatment path when small to intermediate-sized lung nodules are identified. We have combined flow cytometry and machine learning to develop a sputum-based test (CyPath Lung) that can assist physicians in decision-making in such cases. METHODS: Single cell suspensions prepared from induced sputum samples collected over three consecutive days were labeled with a viability dye to exclude dead cells, antibodies to distinguish cell types, and a porphyrin to label cancer-associated cells. The labeled cell suspension was run on a flow cytometer and the data collected. An analysis pipeline combining automated flow cytometry data processing with machine learning was developed to distinguish cancer from non-cancer samples from 150 patients at high risk of whom 28 had lung cancer. Flow data and patient features were evaluated to identify predictors of lung cancer. Random training and test sets were chosen to evaluate predictive variables iteratively until a robust model was identified. The final model was tested on a second, independent group of 32 samples, including six samples from patients diagnosed with lung cancer. RESULTS: Automated analysis combined with machine learning resulted in a predictive model that achieved an area under the ROC curve (AUC) of 0.89 (95% CI 0.83-0.89). The sensitivity and specificity were 82% and 88%, respectively, and the negative and positive predictive values 96% and 61%, respectively. Importantly, the test was 92% sensitive and 87% specific in cases when nodules were < 20 mm (AUC of 0.94; 95% CI 0.89-0.99). Testing of the model on an independent second set of samples showed an AUC of 0.85 (95% CI 0.71-0.98) with an 83% sensitivity, 77% specificity, 95% negative predictive value and 45% positive predictive value. The model is robust to differences in sample processing and disease state. CONCLUSION: CyPath Lung correctly classifies samples as cancer or non-cancer with high accuracy, including from participants at different disease stages and with nodules < 20 mm in diameter. This test is intended for use after lung cancer screening to improve early-stage lung cancer diagnosis. Trial registration ClinicalTrials.gov ID: NCT03457415; March 7, 2018.

Junctional instability in neuroepithelium and network hyperexcitability in a focal cortical dysplasia human model.

Focal cortical dysplasia is a highly epileptogenic cortical malformation with few treatment options. Here, we generated human cortical organoids from patients with focal cortical dysplasia type II. Using this human model, we mimicked some focal cortical dysplasia hallmarks, such as impaired cell proliferation, the presence of dysmorphic neurons and balloon cells, and neuronal network hyperexcitability. Furthermore, we observed alterations in the adherens junctions zonula occludens-1 and partitioning defective 3, reduced polarization of the actin cytoskeleton, and fewer synaptic puncta. Focal cortical dysplasia cortical organoids showed downregulation of the small GTPase RHOA, a finding that was confirmed in brain tissue resected from these patients. Functionally, both spontaneous and optogenetically-evoked electrical activity revealed hyperexcitability and enhanced network connectivity in focal cortical dysplasia organoids. Taken together, our findings suggest a ventricular zone instability in tissue cohesion of neuroepithelial cells, leading to a maturational arrest of progenitors or newborn neurons, which may predispose to cellular and functional immaturity and compromise the formation of neural networks in focal cortical dysplasia.

Long-term Ecological Research: Chasing fashions or being prepared for fashion changes?

Long-term-ecological-research (LTER) faces many challenges, including the difficulty of obtaining long-term funding, changes in research questions and sampling designs, keeping researchers collecting standardized data for many years, impediments to interactions with local people, and the difficulty of integrating the needs of local decision makers with "big science". These issues result in a lack of universally accepted guidelines as to how research should be done and integrated among LTER sites. Here we discuss how the RAPELD (standardized field infrastructure system), can help deal with these issues as a complementary technique in LTER studies, allowing comparisons across landscapes and ecosystems and reducing sampling costs. RAPELD uses local surveys to understand broad spatial and temporal patterns while enhancing decision-making and training of researchers, local indigenous groups and traditional communities. Sampling of ecological data can be carried out by different researchers through standardized protocols, resulting in spatial data that can be used to answer temporal questions, and allow new questions to be investigated. Results can also be integrated into existing biodiversity networks. Integrated systems are the most efficient way to save resources, maximize results, and accumulate information that can be used in the face of the unknown unknowns upon which our future depends.

Current situation of endemic mycosis in the Americas and the Caribbean: Proceedings of the first international meeting on endemic mycoses of the Americas (IMEMA).

BACKGROUND: The Americas are home to biologically and clinically diverse endemic fungi, including Blastomyces, Coccidioides, Emergomyces, Histoplasma, Paracoccidioides and Sporothrix. In endemic areas with high risk of infection, these fungal pathogens represent an important public health problem. OBJECTIVES: This report aims to summarise the main findings of the regional analysis carried out on the status of the endemic mycoses of the Americas, done at the first International Meeting on Endemic Mycoses of the Americas (IMEMA). METHODS: A regional analysis for the Americas was done, the 27 territories were grouped into nine regions. A SWOT analysis was done. RESULTS: All territories reported availability of microscopy. Seventy percent of territories reported antibody testing, 67% of territories reported availability of Histoplasma antigen testing. None of the territories reported the use of (1-3)-ß-d-glucan. Fifty two percent of territories reported the availability of PCR testing in reference centres (mostly for histoplasmosis). Most of the territories reported access to medications such as trimethoprim-sulfamethoxazole, itraconazole, voriconazole and amphotericin B (AMB) deoxycholate. Many countries had limited access to liposomal formulation of AMB and newer azoles, such as posaconazole and isavuconazole. Surveillance of these fungal diseases was minimal. CONCLUSIONS: A consensus emerged among meeting participants, this group concluded that endemic mycoses are neglected diseases, and due to their severity and lack of resources, the improvement of diagnosis, treatment and surveillance is needed.

Assessment of the Oral Health Impact Profile (OHIP-14) improvement of different treatments for dentin hypersensitivity in noncarious cervical lesions-a randomized clinical study.

OBJECTIVES: To assess the improvement on oral health-related quality of life (OHRQoL) of individuals with cervical dentin hypersensitivity (CDH) with the noncarious cervical lesion (NCCLs) subjected to different treatments. MATERIAL AND METHODS: A single-blind randomized clinical trial was conducted with patients (n = 74) randomly allocated into three groups according to the treatment performed: PO, potassium oxalate (Oxa-Gel BF); LL, low-power laser irradiation; and POLL, potassium oxalate (Oxa-Gel BF) associated with low-power laser irradiation. The treatments were applied in four steps, weekly. The questionnaire Oral Health Impact Profile (OHIP-14) was applied before initiating and immediately after the fourth session. The greater the sum of the score, the smaller the measurement of OHRQoL. RESULTS: The total mean values of the variables at the final moment were significantly lower than the initial one for all dimensions (p value ranging from < 0.001 to 0.006). Furthermore, the OHIP-14 scores final analysis between the groups indicated that the POLL group compared to LL had a significantly lower final score for the functional limitation (p = 0.009), physical pain (p = 0.049), and psychological discomfort (p = 0.035) dimensions and that group PO compared to group LL had a significantly lower final score for the functional limitation dimension (p = 0.024). CONCLUSIONS: There was a reduction in the OHIP-14 score for all dimensions, indicating an improvement in patients' quality of life after the use of desensitizing therapies. Patients in group LL had a higher functional limitation, physical pain, and psychological discomfort. CLINICAL RELEVANCE: This study indicated improvement of desensitizing therapies for CDH to improve patients' OHRQoL. TRIAL REGISTRATION: This trial was registered in the Brazilian Clinical Trials Registry Platform (REBEC protocol number RBR-4ybjmt).

Health-related quality of life predictors in people with chronic wounds.

OBJECTIVE: To assess health-related quality of life and its predictors among people with chronic wounds. MATERIAL AND METHODS: A cross-sectional, analytical study was conducted with 85 people with chronic wounds, from July to September 2019, at a university hospital in the state of Paraíba, Brazil. Data were collected using forms and the Cardiff Wound Impact Schedule (CWIS). For data analysis, multiple linear regression was performed. RESULTS: The mean quality of life score according to the CWIS was 45.57. The variables gender, marital status, comorbidities, duration of injury and appearance of exudate were negatively associated with the well-being domain. Injury duration, tissue depth, tissue type, and exudate volume were variables negatively associated with physical symptoms and mastery of daily living. Religion, wound origin, duration, lesion size, and tissue depth were negatively associated with the social life domain. CONCLUSION: Sociodemographic characteristics and wound-related aspects need to be considered when assessing and caring for patients with chronic wounds to ensure a better quality of life.

In vivo study of hypericin-loaded poloxamer-based mucoadhesive in situ gelling liquid crystalline precursor system in a mice model of vulvovaginal candidiasis.

The present study reports the performance of the pigment hypericin (HYP)-loaded poloxamer-based mucoadhesive in situ gelling liquid crystalline precursor system (LCPS) for the treatment of vulvovaginal candidiasis (VVC) in mice. LCPS composed of 40% of ethoxylated and propoxylated cetyl alcohol, 30% of oleic acid and cholesterol (7:1), 30% of a dispersion of 16% poloxamer 407 and 0.05% of HYP (HYP-LCPS) was prepared and characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and ex vivo permeation and retention studies across vaginal porcine mucosa were performed. In addition, the antifungal properties of the HYP-LCPS were evaluated in a murine in vivo model; for this, infected C57BL female mice groups were treated with both HYP in solution and HYP-LCPS, and after 6 days colony forming unit (CFU)/ml count was performed. PLM and SAXS confirmed that HYP-LCPS is a microemulsion situated in boundary transition region confirming its action as an LCPS. When in contact with simulated vaginal fluid, HYP-LCPS became rigid and exhibited maltase crosses and bragg peaks characteristics of lamellar phase. Ex vivo permeation and retention studies showed that HYP-LCPS provides a localized treatment on the superficial layers of porcine vaginal mucosa. HYP-LCPS induced a significant reduction in the number of CFU/ml in the mice; thus this formulation indicated it is as effective as a commercial dosage form. It was concluded that LCPS maintains the biological activity of HYP and provides an adequate drug delivery system for this lipophilic molecule at the vaginal mucosa, being a promising option in cases of VVC.

Pesticides detected in surface and groundwater from agroecosystems in the Pampas region of Argentina: occurrence and ecological risk assessment.

The objective of this study was to evaluate the occurrence of pesticides in surface and groundwater of agricultural areas of the Pampas region of Argentina and to develop an ecological risk assessment (ERA) of pesticides in freshwater ecosystems. Eight agricultural sites from south Santa Fe province, in the north of the Pampas region, were sampled seven times between 2016 and 2018. Pesticides were analysed by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry (UPLC-LC/MS). Twenty compounds among herbicides, insecticides and fungicides in 84% and 79% of groundwater and surface water samples, respectively, were detected. Atrazine was the most ubiquitous pesticide, following by metolachlor, acetochlor and glyphosate, with maximum concentrations of 28, 24, 77 and 111 µg/L, respectively. An ERA was performed by employing the risk quotient (RQ) method. Atrazine, azoxystrobin, pirimiphos-methyl, acetochlor and epoxiconazole posed a high and very high risk for aquatic organisms (RQ > 1) and glyphosate, metolachlor and 2,4-D exhibited negligible to medium risk. The herbicides were the major contributors to risk. This study is the first contribution on the presence and concentration of pesticides in surface and groundwater from agricultural areas of south Santa Fe province, north Pampas region, Argentina, and a starting point for pesticide ecological risk assessment.

[Identification and susceptibility profile of Candida species isolated from hemocultures in hospitals in ParaguayIdentificação e perfil de sensibilidade de Candida spp. isoladas de hemoculturas em hospitais no Paraguai]. / Identificación y perfil de sensibilidad de Candida spp. aisladas de hemocultivos en hospitales de Paraguay.

OBJECTIVE: To report the species identified in candidemia isolated in hospitals in Paraguay, their distribution by age groups, and their susceptibility to fluconazole, voriconazole, and amphotericin B. METHODS: Retrospective study of candidemia reported to the Central Health Public Laboratory by seven hospitals between 2010 and 2018. Identification and susceptibility testing were performed using the automated Vitek 2® system. RESULTS: 520 cases of candidiasis were reported. The prevalent species were Candida albicans (34.4%), C. parapsilosis (30.4%), C. tropical (25.4%), C. glabrata (4.8%), and C. krusei (2.1%). Less frequent species were: C. lusitaniae (1.2%), C. guilliermondii (1%), C. famata (0.3%), C. rugous (0.2%), and C. kefyr (0.2%). Candidiasis was most prevalent in the elderly (48.5%) and adults (29.9%). C. parapsilosis was most frequent in newborns and infants, and C. tropical in pediatric patients; 2.8% and 0.6% of C. albicans and 4.4% and 2.5% of C. parapsilosis were resistant, respectively, to fluconazole and voriconazole; 8% of C. glabrata were resistant to fluconazole and two isolations showed a minimum inhibitory concentration for voriconazole greater than 0.25 mg/L. CONCLUSIONS: This is the first report on candidemias in hospitals of Paraguay. Together with other studies, it will improve knowledge about this infection in the country.

OBJETIVO: Relatar as espécies identificadas em candidemias isoladas em hospitais do país, apresentar a sua distribuição por faixa etária e determinar a sua sensibilidade ao fluconazol, voriconazol e anfotericina B. MÉTODOS: Estudo retrospectivo de candidemias notificadas por sete hospitais ao Laboratório Central de Saúde Pública no período de 2010 a 2018. A identificação e o antifungigrama foram realizados pelo sistema automatizado Vitek®2. RESULTADOS: Foram notificados 520 casos de candidemia. As espécies predominantes foram Candida albicans (34,4%), C. parapsilosis (30,4%), C. tropicalis (25,4%), C. glabrata (4,8%) e C. krusei (2,1%). Outras espécies menos frequentes foram: C. lusitaniae (1,2%), C. guilliermondii (1%), C. famata (0,3%), C. rugosa (0,2%) e C. kefyr (0,2%). As candidemias em pacientes idosos (48,5%) e adultos (29,9%) foram as mais prevalentes. C. parapsilosis foi a espécie mais frequente em recém-nascidos e lactentes, e C. tropicalis predominou em pacientes pediátricos. Ao todo, 2,8% e 0,6% dos casos de C. albicans e 4,4% e 2,5% dos de C. parapsilosis foram resistentes a fluconazol e voriconazol, respectivamente. Além disso, 8% dos casos de C. glabrata foram resistentes ao fluconazol, e duas amostras isoladas apresentaram uma concentração inibitória mínima para voriconazol acima de 0,25 mg/L. CONCLUSÕES: Este é o primeiro estudo sobre candidemias em hospitais do Paraguai, o que, juntamente com outros estudos, permitirá conhecer melhor esta infecção no país.

Dysregulation of NEUROG2 plays a key role in focal cortical dysplasia.

OBJECTIVE: Focal cortical dysplasias (FCDs) are an important cause of drug-resistant epilepsy. In this work, we aimed to investigate whether abnormal gene regulation, mediated by microRNA, could be involved in FCD type II. METHODS: We used total RNA from the brain tissue of 16 patients with FCD type II and 28 controls. MicroRNA expression was initially assessed by microarray. Quantitative polymerase chain reaction, in situ hybridization, luciferase reporter assays, and deep sequencing for genes in the mTOR pathway were performed to validate and further explore our initial study. RESULTS: hsa-let-7f (p = 0.039), hsa-miR-31 (p = 0.0078), and hsa-miR34a (p = 0.021) were downregulated in FCD type II, whereas a transcription factor involved in neuronal and glial fate specification, NEUROG2 (p < 0.05), was upregulated. We also found that the RND2 gene, a NEUROG2-target, is upregulated (p < 0.001). In vitro experiments showed that hsa-miR-34a downregulates NEUROG2 by binding to its 5'-untranslated region. Moreover, we observed strong nuclear expression of NEUROG2 in balloon cells and dysmorphic neurons and found that 28.5% of our patients presented brain somatic mutations in genes of the mTOR pathway. INTERPRETATION: Our findings suggest a new molecular mechanism, in which NEUROG2 has a pivotal and central role in the pathogenesis of FCD type II. In this way, we found that the downregulation of hsa-miR-34a leads to upregulation of NEUROG2, and consequently to overexpression of the RND2 gene. These findings indicate that a faulty coupling in neuronal differentiation and migration mechanisms may explain the presence of aberrant cells and complete dyslamination in FCD type II. Ann Neurol 2018;83:623-635.

Mucoadhesive In Situ Gelling Liquid Crystalline Precursor System to Improve the Vaginal Administration of Drugs.

The vaginal mucosa is a very promising route for drug administration due to its high permeability and the possibility to bypass first pass metabolism; however, current vaginal dosage forms present low retention times due to their dilution in vaginal fluids, which hampers the efficacy of many pharmacological treatments. In order to overcome these problems, this study proposes to develop a mucoadhesive in situ gelling liquid crystalline precursor system composed of 30% of oleic acid and cholesterol (7:1), 40% of ethoxylated and propoxylated cetyl alcohol, and 30% of a dispersion of 16% Poloxamer 407. The effect of the dilution with simulated vaginal fluid (SVF) on this system was evaluated by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), rheological studies, texture profile analysis (TPA), mucoadhesion study, in vitro drug release test using hypericin (HYP) as drug model, and cytotoxicity assay. PLM and SAXS confirmed the formation of an isotropic system. After the addition of three different concentrations of SVF (30, 50, and 100%), the resultant formulations presented anisotropy and characteristics of viscous lamellar phases. Rheology shows that formulations with SVF behaved as a non-Newtonian fluid with suitable shear thinning for vaginal application. TPA and mucoadhesion assays indicated the formation of long-range ordered systems as the amount of SVF increases which may assist in the fixation of the formulation on the vaginal mucosa. The formulations were able to control about 75% of the released HYP demonstrating a sustained release profile. Finally, all formulations acted as safe vaginal drug delivery systems.

Mechanical Complications Related to the Retention Screws of Prefabricated Metal Abutments With Different Angulations: A Retrospective Study With 916 Implants.

OBJECTIVES: The present retrospective study assessed the clinical performance of abutment screws from prefabricated metal abutments and compared technical complication rates between straight and angled abutments. MATERIALS AND METHODS: Dental charts were selected for patients with dental implant rehabilitations delivered between 1998 and 2012. Abutment angulation, prosthetic screw type, and presence of complications that occurred during the selected time period were collected. Technical complications registered included abutment screw loosening and/or fractures detected during clinical and radiographic examinations. The chi-square test was used for statistical analysis. RESULTS: Abutment angulations were divided into 2 groups: G1) prefabricated straight abutments and G2) prefabricated angled conical mini UCLA-type abutments. A total of 916 implants (799 straight and 117 angled conical mini UCLA-type abutments) were evaluated. G1 showed 91.1% had absence of failures, which were clinically defined as any screw loosening or fracture; and 8.9% reported some type of technical complication. G2 showed 92.3% and 7.7%, with and without technical complications, respectively. CONCLUSIONS: No significant differences were observed between abutment angulation and technical complications.

Musashi1 Impacts Radio-Resistance in Glioblastoma by Controlling DNA-Protein Kinase Catalytic Subunit.

The conserved RNA-binding protein Musashi1 (MSI1) has been characterized as a stem cell marker, controlling the balance between self-renewal and differentiation and as a key oncogenic factor in numerous solid tumors, including glioblastoma. To explore the potential use of MSI1 targeting in therapy, we studied MSI1 in the context of radiation sensitivity. Knockdown of MSI1 led to a decrease in cell survival and an increase in DNA damage compared to control in cells treated with ionizing radiation. We subsequently examined mechanisms of double-strand break repair and found that loss of MSI1 reduces the frequency of nonhomologous end-joining. This phenomenon could be attributed to the decreased expression of DNA-protein kinase catalytic subunit, which we have previously identified as a target of MSI1. Collectively, our results suggest a role for MSI1 in double-strand break repair and that its inhibition may enhance the effect of radiotherapy.

High-throughput analyses of hnRNP H1 dissects its multi-functional aspect.

hnRNPs are polyvalent RNA binding proteins that have been implicated in a range of regulatory roles including splicing, mRNA decay, translation, and miRNA metabolism. A variety of genome wide studies have taken advantage of methods like CLIP and RIP to identify the targets and binding sites of RNA binding proteins. However, due to the complex nature of RNA-binding proteins, these studies are incomplete without assays that characterize the impact of RBP binding on mRNA target expression. Here we used a suite of high-throughput approaches (RIP-Seq, iCLIP, RNA-Seq and shotgun proteomics) to provide a comprehensive view of hnRNP H1s ensemble of targets and its role in splicing, mRNA decay, and translation. The combination of RIP-Seq and iCLIP allowed us to identify a set of 1,086 high confidence target transcripts. Binding site motif analysis of these targets suggests the TGGG tetramer as a prevalent component of hnRNP H1 binding motif, with particular enrichment around intronic hnRNP H1 sites. Our analysis of the target transcripts and binding sites indicates that hnRNP H1s involvement in splicing is 2-fold: it directly affects a substantial number of splicing events, but also regulates the expression of major components of the splicing machinery and other RBPs with known roles in splicing regulation. The identified mRNA targets displayed function enrichment in MAPK signaling and ubiquitin mediated proteolysis, which might be main routes by which hnRNP H1 promotes tumorigenesis.

Nucleolar proteins regulate stage-specific gene expression and ribosomal RNA maturation in Trypanosoma brucei.

Different life-cycle stages of Trypanosoma brucei are characterized by stage-specific glycoprotein coats. GPEET procyclin, the major surface protein of early procyclic (insect midgut) forms, is transcribed in the nucleolus by RNA polymerase I as part of a polycistronic precursor that is processed to monocistronic mRNAs. In culture, when differentiation to late procyclic forms is triggered by removal of glycerol, the precursor is still transcribed, but accumulation of GPEET mRNA is prevented by a glycerol-responsive element in the 3' UTR. A genome-wide RNAi screen for persistent expression of GPEET in glycerol-free medium identified a novel protein, NRG1 (Nucleolar Regulator of GPEET 1), as a negative regulator. NRG1 associates with GPEET mRNA and with several nucleolar proteins. These include two PUF proteins, TbPUF7 and TbPUF10, and BOP1, a protein required for rRNA processing in other organisms. RNAi against each of these components prolonged or even increased GPEET expression in the absence of glycerol as well as causing a significant reduction in 5.8S rRNA and its immediate precursor. These results indicate that components of a complex used for rRNA maturation can have an additional role in regulating mRNAs that originate in the nucleolus.

Knowledge of the workers of the construction industry about acquired immunodeficiency syndrome and associated factors.

BACKGROUND: Sexually Transmitted Infections (STIs), such as Human Immunodeficiency Virus (HIV) transmitted by biological, socioeconomic and cultural factors that increase the vulnerability of certain population groups such as civil construction workers. OBJECTIVE: To evaluate the knowledge of workers in the construction industry about infection by HIV and associated factors. METHODS: A cross-sectional, analytical study was conducted, including 381 construction workers, in the cities of João Pessoa, Cabedelo, and Santa Rita, in the State of Paraíba, Brazil, performed through interviews, using a data collection instrument with social demographic variables and the HIV Knowledge Questionnaire. Data were analyzed using descriptive measures, and for comparison between the categories the Kruskal-Wallis test was used, with margin of error of 5% . RESULTS: A predominance of male workers was observed (97.4%) among 381 workers; aged 30-39 (34.1%); married (81.4%); with up to eight years of education (66.6%); family income of up to 2x the minimum wage (52.8%) and coming from the country (43.8%). The average of correct responses was 63.3%, with higher indices (70.0%) related to transmission and forms of prevention. There were higher percentages of hits among those who had studied for 12 years or more. Workers from the capital of Paraíba presented a higher number of correct responses (67.44%). CONCLUSIONS: There is a low index of knowledge of construction workers about HIV, highlighting their consequent vulnerability to acquiring this infection.

Distinct genomic organization, mRNA expression and cellular localization of members of two amastin sub-families present in Trypanosoma cruzi.

BACKGROUND: Amastins are surface glycoproteins (approximately 180 residues long) initially described in Trypanosoma cruzi as particularly abundant during the amastigote stage of this protozoan parasite. Subsequently, they have been found to be encoded by large gene families also present in the genomes of several species of Leishmania and in other Trypanosomatids. Although most amastin genes are organized in clusters associated with tuzin genes and are up-regulated in the intracellular stage of T. cruzi and Leishmania spp, distinct genomic organizations and mRNA expression patterns have also been reported. RESULTS: Based on the analysis of the complete genome sequences of two T. cruzi strains, we identified a total of 14 copies of amastin genes in T. cruzi and showed that they belong to two of the four previously described amastin subfamilies. Whereas δ-amastin genes are organized in two or more clusters with alternating copies of tuzin genes, the two copies of ß-amastins are linked together in a distinct chromosome. Most T. cruzi amastins have similar surface localization as determined by confocal microscopy and western blot analyses. Transcript levels for δ-amastins were found to be up-regulated in amastigotes from several T. cruzi strains, except in the G strain, which is known to have low infection capacity. In contrast, in all strains analysed, ß-amastin transcripts are more abundant in epimastigotes, the stage found in the insect vector. CONCLUSIONS: Here we showed that not only the number and diversity of T. cruzi amastin genes is larger than what has been predicted, but also their mode of expression during the parasite life cycle is more complex. Although most T. cruzi amastins have a similar surface localization, only δ-amastin genes have their expression up-regulated in amastigotes. The results showing that a sub-group of this family is up-regulated in epimastigotes, suggest that, in addition of their role in intracellular amastigotes, T. cruzi amastins may also serve important functions during the insect stage of the parasite life cycle. Most importantly, evidence for their role as virulence factors was also unveiled from the data showing that δ-amastin expression is down regulated in a strain presenting low infection capacity.