Predictors of dropout in the school-based multi-component intervention, 'Mexa-se'.

To identify the predictors of dropout in the 'Mexa-se' intervention according to the body mass index (BMI) category. This was a controlled, non-randomized study. The intervention included: (i) increase in the intensity of physical activities (PA) in physical education (PE) classes; (ii) active recess; (iii) educational sessions on PA, nutrition and body image; and (iv) educational materials. Dropout was considered when students dropped out of intervention, or did not reach 75% attendance in PE classes. The independent variables were gender, age, study period, socioeconomic status, BMI, PA, screen time, food consumption, health perception, attitudes toward PA, self-efficacy for PA, perception of the school environment, body image and self-esteem. Binary logistic regression analysis was used. The dropout rate was 26.8%. In the total sample and among students with an adequate BMI, there was a greater probability of dropout with an increase in age. For overweight students, increased age and socioeconomic status, and studying in the afternoon period were predictors of dropout from the intervention. Socio-demographic factors were predictors of dropout from the 'Mexa-se' intervention; the associated factors differed based on the BMI category.

Multifunctional laminarin microparticles for cell adhesion and expansion.

Microfabrication technologies have been widely explored to produce microgels that can be assembled in functional constructs for tissue engineering and regenerative medicine applications. Here, we propose microfluidics coupled to a source of UV light to produce multifunctional methacrylated laminarin microparticles with narrow distribution of sizes using photopolymerization. The multifunctional microparticles were loaded with platelet lysates and further conjugated with an adhesive peptide. The adhesive peptides dictated cell adhesiveness to the laminarin microparticles, the incorporation of platelet lysates have resulted in improved cell expansion compared to clear microparticles. Overall, our findings demonstrate that multifunctional methacrylated laminarin microparticles provide an effective support for cell attachment and expansion. Moreover, expanded cells provide the link for microparticles aggregation resulting in robust 3D structures. This suggest the potential for using the methacrylated laminarin microplatforms capable to be assembled by the action of cells to rapidly produce large tissue engineered constructs.

Determination of micronucleus frequency by acridine orange fluorescent staining in peripheral blood reticulocytes of mice treated topically with different lubricant oils and cyclophosphamide.

To ascertain whether used and re-refined lubricant oil absorbed through the skin can produce a genotoxic effect or cytotoxicity in mouse bone marrow cells, we examined the induction of micronucleated erythrocytes of peripheral blood after cutaneous application. Both re-refined and used lubricant oils showed a weak but significant induction of micronucleated polychromatic erythrocytes compared with control, while virgin oil did not show micronucleus induction. Cyclophosphamide (CP) was used not only as positive control but also to compare the sensitivity between intraperitoneal and dermal routes of administration of the test compounds in mice. The efficacy of intraperitoneal injection of CP is well known. On the other hand, dermal exposure is not so common and when CP was diluted in glycerin statistically significant values (P = 0.0036) of micronuclei were also found. Topically applied lubricant oils (virgin, re-refined and used) have the capacity to interfere with mouse bone marrow hematopoiesis evidenced by a statistically significant decrease in the proportion of polychromatic erythrocytes in the peripheral blood. Physical and chemical analysis revealed that used oil is more viscous than other lubricants, suggesting the presence of insoluble compounds, oxidized products and water as well as aromatic hydrocarbons. Used oil differs from other lubricant oils in metal and polyaromatic hydrocarbon content. Re-refined oil revealed a neutral value typical of pure mineral oil. This assay is an important tool to evaluate environmental pollutants that cause genotoxicity and/or cytotoxicity through skin exposure.

L1 sequence of a new human papillomavirus type-58 variant associated with cervical intraepithelial neoplasia.

The present study on molecular characterization of a human papillomavirus (HPV) isolated in Central Brazil describes the L1 gene sequence from a new variant of HPV-58, the isolate Bsb-02. The sample was from a smear obtained from a woman with cervical intraepithelial neoplasia grade II. The whole L1 gene from isolate Bsb-02 was sequenced automatically, showing 99.1% nucleotide identity with the gene from the HPV-58 reference. The clustering between Bsb-02 and HPV-58 reference sequence was also supported by phylogenetic analysis. Fourteen nucleotide substitutions were observed: eight were synonymous and six were associated with amino acid substitutions. A10V and V144I have not been previously described. At GenBank, the only complete L1 sequence from HPV-58 in addition to the HPV-58 reference one is that of Bsb-02. These data provide information that may be relevant to HPV diagnosis and to rational vaccine strategies. HPV variants may also be associated with host immune responses and with the risk of cervical neoplasia.

Characterization of pemphigus foliaceus antigen from human epidermis.

Pemphigus foliaceus (PF) and its endemic form, Fogo Selvagem (FS), are characterized by subcorneal vesicles and pathogenic IgG autoantibodies directed against keratinocyte surface antigens. A major pool of FS antigen(s) remains bound to the insoluble epidermal envelope fraction. In this paper we demonstrate that this antigen(s) can be released from the envelope fraction by sonication. By immune precipitation four components can be detected, having molecular weights (MW) of 260, 80, 62, and 45 kD. The 260-kD component is lost by boiling or extraction with glycine HCl at pH 2.8. The major components appear to be the 80- and 62-kD poly-peptides. They chromatograph as a unit by gel filtration in 0.1% SDS, in the MW range of 115-120 kD. The FS antigen(s) appears to be cationic, forming insoluble complexes at low pH with SDS, and is labile to ammonium sulfate and freezing and thawing. It is unaffected by positive pressure concentration, 50% acetone precipitation, and reduction/alkylation. The FS antigen(s) is precipitated by all FS and nonendemic PF sera except those in complete clinical and serologic remission. The FS antigen(s) is also precipitated by 50% of pemphigus vulgaris but none of the bullous pemphigoid sera tested. All FS antigenic components are immunoprecipitated by IgG4 autoantibodies, but the IgG1 subclass from the same patients appear to immunoprecipitate only the 62-kD polypeptide. The FS antigen(s) is able to adsorb human autoantibodies against human desmoglein 1 (DG1), but not rabbit antisera against bovine DG1 or 2. This paper shows that physical stress, i.e., sonication, may be able to solubilize sufficient FS antigen(s) from the epidermal envelope fractions for further chemical characterization. The relationship of these FS antigen(s) to other reported FS antigens is presently unknown.

A soluble and immunoreactive fragment of pemphigus foliaceus antigen released by trypsinization of viable human epidermis.

Pemphigus foliaceus (PF) antigen is a transmembrane desmosomal glycoprotein (desmoglein I), part of which is located on the keratinocyte surface. Previous studies have shown that after trypsinization of viable human epidermis, this antigen is no longer detected on the surface of detached keratinocytes. It was not known, however, if this loss of antigenic activity was due to destruction, internalization, or cleavage of the antigen itself. In the present study we investigated the fate of the PF antigen after trypsinization of viable human skin. By using Concanavalin-A agarose affinity chromatography, we could partially purify an antigenic glycoprotein fraction that was released by trypsinization into the medium. This antigenic fraction was radiolabeled and tested by immunoprecipitation using sera from endemic pemphigus foliaceus or fogo selvagem (FS), non-endemic pemphigus foliaceus (NEPF), pemphigus vulgaris (PV), and bullous pemphigoid (BP) patients, and sera from normal subjects as controls. Immunoprecipitated labeled proteins were analyzed by SDS-PAGE and autoradiography. All FS sera (20 of 20 FS and five of five NEPF) and 46% of the PV sera (six of 13) immunoprecipitated a band of 45-kD molecular weight. Sera from FS patients in prolonged clinical and serological remission (seven of 10), sera from BP patients (five of five), and sera from normal donors (nine of nine) did not precipitate this 45-kD band. This study showed that a fragment of the PF antigen is released by trypsinization of human skin as a soluble immunoreactive glycopeptide of 45-kD molecular weight. Additionally, this procedure has generated sufficient quantities of the PF antigen for further biochemical characterization.

Serologic abnormalities in patients with endemic pemphigus foliaceus (Fogo selvagem), their relatives, and normal donors from endemic and non-endemic areas of Brazil.

Based on epidemiologic data, a current hypothesis states that Fogo selvagem (FS) may be triggered by environmental factors present in endemic areas of Brazil. Because the appearance of new cases is limited to those areas, we wanted to ascertain if the presence of the pemphigus autoantibodies was restricted to the patients. To further delineate the restriction of the autoantibody response in these patients we also investigated the presence of lupus-associated autoantibodies. Using indirect immunofluorescence (IF) we tested the sera of patients with FS (n = 196), their relatives (n = 138), their cohabitants (n = 13), and normal donors from endemic (n = 38) and non-endemic areas (n = 44) for pemphigus autoantibodies. Antinuclear antibodies (ANA) and anti-nDNA antibodies were determined by indirect IF against Hep-2 cells and Crithidia lucilliae, respectively. Autoantibodies against nRNP, Ro/SSA, La/SSB, and Sm were assayed by double immune diffusion in agarose gels. FS autoantibodies were present in the sera of all patients with active disease (n = 196, 100%, titers greater than 40 to 2560), but were not found in any sera from normal individuals in endemic or non-endemic areas. The titer of the FS autoantibody showed a rough correlation with the extent and activity of the disease. Furthermore, lupus-associated autoantibodies were not present in any of the tested samples. We conclude the FS antiepidermal autoantibodies are specific serologic markers of the disease and are not present in unaffected individual from the endemic areas. As such, they provide an important marker that should be useful in ongoing epidemiologic studies aimed at identifying putative etiologic agent(s).

Pemphigus foliaceus antigen: characterization of a keratinocyte envelope associated pool and preparation of a soluble immunoreactive fragment.

In both the endemic and sporadic forms of pemphigus foliaceus (PF), antiepidermal autoantibodies against desmoglein I are present. Desmoglein I is a highly insoluble 160-kD transmembrane glycoprotein of the desmosomal core. The detailed immunochemical characterization of the epitope(s) recognized by the PF autoantibodies is hampered by its large molecular weight and the insolubility of desmoglein I in nondenaturing buffers. This study was designed to identify alternative methods that could yield soluble immunoreactive PF antigen (Ag) from normal human epidermis. The presence of PF Ag in human epidermis and in its soluble or insoluble fractions was monitored by indirect immunofluorescence, immunoadsorption of PF sera, and immunoprecipitation of radiolabeled fractions. The PF Ag from trypsin-resistant, radiolabeled cell envelope preparations was cleaved by papain and immunoprecipitated by PF sera. A 50-kD peptide, isoelectric at pH 5.5-5.8, was immunoprecipitated by sera from all patients with endemic PF (n = 15) or idiopathic PF (n = 4), and by two of four pemphigus vulgaris sera, but by no control sera (n = 7). This study shows that a significant fraction of the PF Ag is insoluble, trypsin-resistant, and is associated with the cornified cell envelope fraction, but an Ag fragment can be obtained in a small molecular weight, soluble, and immunoreactive form by papain digestion. This 50-kD papain fragment is more amenable to detailed chemical and immunologic characterization than the native molecule.

Calcium enhances the sensitivity of immunofluorescence for pemphigus antibodies.

Indirect immunofluorescence (IF) to detect pemphigus and pemphigoid autoantibodies is commonly performed with monkey esophagus (ME) as substrate and phosphate-buffered saline (PBS) as a diluent. The purpose of this study was to evaluate comparative IF titers using human skin (HS) as substrate with variations in the buffers employed. Substrates (ME or HS) were incubated in PBS, Tris-acetate-buffered saline (TAS), TAS with 5 mM CaCl+2 (TAS-Ca+2), and PBS or TAS with 1 mM EDTA, prior to incubation with pemphigus or pemphigoid sera for indirect IF. We examined sera from 11 patients with pemphigus vulgaris (PV), 10 patients with Brazilian pemphigus foliaceus (BPF), and 4 patients with bullous pemphigoid. In 20 of 21 pemphigus sera, endpoint indirect IF titers were highest on normal skin with TAS-Ca+2. Six sera (2 PV and 4 BPF) had endpoints that were 5 double dilutions higher than the endpoints obtained with ME and PBS. Six sera (3 PV and 3 BPF) were 4 double dilutions higher, 7 sera (3 PV and 4 BPF) were 2-3 double dilutions higher, and 2 PV sera were equivalent with both substrate/buffers. Preincubation of either tissue with EDTA prior to indirect IF abolished PV and BPF antibody binding completely. Exposure to EDTA after the tissue was incubated with PV or BPF sera did not affect indirect IF titers. In the presence of Ca+2, the antigen was resistant to trypsin in concentrations of 0.001%; however, in the absence of added Ca+2 it was destroyed by 0.0001% trypsin. These differences were not observed with bullous pemphigoid sera; all 4 sera had similar endpoint indirect IF titers. This study shows a significant increase in the sensitivity of indirect IF assays for pemphigus autoantibodies by the use of Ca+2-supplemented buffers on human skin. This finding may also have implications for procedures designed to purify and/or detect pemphigus antigens.

Environmental risk factors in endemic pemphigus foliaceus (Fogo selvagem). "The Cooperative Group on Fogo Selvagem Research".

Endemic pemphigus foliaceus or Fogo selvagem (FS) is an epidermal organ-specific autoimmune disease mediated by autoantibodies. Individuals at risk are peasants who live and work on farms located in the interior of certain endemic states of Brazil. This case-control study compares a group of 52 FS patients with 52 patients suffering from other dermatoses admitted and followed at the hospital for pemphigus (Hospital do Penfigo) in the city of Goiania, state of Goias. Patients and controls matched 1:1 by age, sex, and occupation were examined by two dermatologists at the time of admission and asked to respond to a prepared questionnaire. This questionnaire concerned current and past (1 and 5 years) exposure to environmental risk factors. The following risk factors were assessed: black fly bites, presence of rodents at home, exposure to cereal dust, exposure to fumes or dust released by tree and shrub removal, and exposure to insecticides. Relative risks were estimated from tabulated data by the odds ratio and tested for significance by the chi-square test. The 95% confidence interval for the odds ratio was also calculated for each of the risk factors. The only risk factor showing an odds ratio significantly different from one was exposure to simuliidae bites (odds ratio 4.7, p less than 0.001). This study reinforces the hypothesis that chronic exposure to black fly antigens may precipitate IgG4 antibody formation in predisposed individuals. These antibodies in turn may cross-react with epidermal antigens and cause acantholysis and the clinical expression of the disease known as FS.

An autoantibody in pemphigus serum, specific for the 59 kD keratin, selectively binds the surface of keratinocytes: evidence for an extracellular keratin domain.

We have identified a novel IgG antikeratin autoantibody in the serum of a Brazilian pemphigus foliaceus patient (Cascas-42). This antibody is specific for the 59 kD acidic murine keratin and its 56.5 kD human counterpart (Moll's catalogue #10), and is distinct from the pemphigus antibody system. Antikeratin autoantibodies present in the Cascas-42 serum were purified by affinity chromatography with a 59 kD murine keratin-agarose column (IAP-Cascas-42 antibodies). The specificity of the IAP-Cascas-42 antibodies was tested by indirect immunofluorescence and immunoelectron microscopy against epidermal cryosections, trypsin-dissociated keratinocytes, and epidermal cell cultures. The serum was also tested with extracts from unlabeled and surface 125I-labeled keratinocytes (Iodo-Gen method) by immunoblot analysis of one- and two-dimensional polyacrylamide gel electrophoresis. The IAP-Cascas-42 antibodies bind the intercellular spaces of murine epidermis, and the cell surfaces of viable, dissociated murine keratinocytes, as well as murine epidermal cells in culture by immunofluorescence and immunoelectron microscopy. These autoantibodies did not stain cytoplasmic keratins and did not react with parallel human epidermal substrates. The Cascas-42 serum identified the 59 kD murine acidic keratin and its 56.5 kD human counterpart in epidermal extracts by two-dimensional polyacrylamide gel electrophoresis and immunoblot analysis. In addition, surface radioiodination of viable murine keratinocytes selectively labeled the 59 kD keratin suggesting that a domain of this molecule is exposed on the cell surface. The 125I-labeled 59 kD keratin was also recognized by the Cascas-42 serum by immunoblotting and autoradiography. These studies suggest that in murine epidermis, the 59 kD keratin is a transmembrane protein with an extracellular domain recognized by the IAP-Cascas-42 antibodies.

Endemic pemphigus foliaceus (Fogo Selvagem): II. Current and historic epidemiologic studies.

This paper details current and historic epidemiologic features of Fogo Selvagem (Endemic pemphigus foliaceus) in Brazil. The following features are described. a) The disease occurs in endemic fashion in regions of Brazil within the states of Goias, Mato Grosso do Sul, Parana, Sao Paulo, and Minas Gerais. It appears that the disease is spreading toward the northwest and west, involving the states of Mato Grosso, Para, Maranhao, Rondonia, Acre, and Amazonas. b) People at risk are young peasants or children of either sex or any race exposed to the local ecology in rural areas of endemic states. Although the disease has been described in urban centers, these occurrences are rare. c) Fogo Selvagem commonly appears in wild areas being colonized and disappears as these areas become urbanized. d) The majority of patients live in close proximity to rivers and within the 10-15 Km flying range of mosquitos or black flies (such as Simulium). It is hypothesized that a black fly, Simulium pruinosum may be the vector that precipitates the disease. f) There is a significant number of Fogo Selvagem in family units where multiple, genetically related individuals are affected. g) Finally, autoantibodies against lupus-associated antigens are not present in the sera of patients with Fogo Selvagem. Clinical examination of the skin, and serologic screening for pemphigus autoantibodies are specific parameters that can be used in the search for the etiologic agents that lead to autoimmune disease of the skin. To identify and prove an etiologic agent for this well-characterized autoimmune disease would be of tremendous importance to the understanding of autoimmune skin diseases, and potentially other organ-specific autoimmune disorders.

Effects of melatonin on orofacial movements in rats.

RATIONALE: While reserpine-induced oral movements (OM), an animal model of tardive dyskinesia, are more persistent in old than in adult rats, old animals present spontaneous OM, which are phenomenologically similar to those presented by reserpine-treated adult rats. We postulate that these OM may be the result of oxidative stress induced by both age and reserpine treatment. OBJECTIVES: We intended to determine the preventative effects of exogenous melatonin (one of the most important endogenous antioxidants) as well as suppression of endogenous melatonin via continuous exposure to light on reserpine- or age-induced OM in rats. METHODS: Adult (4 months of age) male Wistar rats were repeatedly treated with saline or melatonin (5 mg/kg, IP) and saline or reserpine and kept under a 12-h light/dark cycle for quantification of reserpine-induced OM as well as oxidative stress (via quantification of lipid peroxidation). To verify the effects of endogenous melatonin suppression on reserpine-induced OM, adult rats were repeatedly treated with saline or reserpine and continuously exposed to light. To verify the effects of exogenous melatonin on age-induced OM older (20 months of age) rats were long-term treated with saline or melatonin and kept under a 12-h light/dark cycle. RESULTS: Melatonin attenuated both reserpine- and age-induced OM. Reserpine enhanced striatal lipid peroxidation, that was prevented by melatonin co-administration. Continuous exposure to light increased spontaneous as well as reserpine-induced OM, indicating that endogenous melatonin may be involved in this movement disorder. CONCLUSIONS: We suggested that melatonin attenuates both reserpine- and age-induced OM in rats.

Study of tooth loss in an adolescent Brazilian population.

Data concerning tooth loss in developing countries may indicate the dental health status in young people and serve as baseline data for evaluation of future dental health programs. The study population consisted of 304 schoolchildren (145 males, 159 females) from Belo Horizonte, M.G. Brazil. The mean age was 14.5, ranging from 13 to 16 yr. The number of teeth already lost and teeth indicated for extraction were assessed from two posterior bitewing radiographs and one frontal color photo which were obtained from all participants. The results showed that 2/3 of the studied population had lost one or more permanent teeth. The average number of missing teeth was 1.8 in both males and females. Of the various teeth, the mandibular first molars had most frequently been lost. Differences between the sexes concerning amount and pattern of lost teeth were small.

Multiple syringomas on the abdomen, thighs, and groin.

Syringomas are benign adnexal tumors that occur most commonly in women. They typically present as soft, flesh-colored to slightly yellow papules on the lower eyelids. We present an unusual case of a healthy 33-year-old male with multiple, reddish brown syringomas located on the lower abdomen, thighs, and groin. Although these lesions can result in significant cosmetic disfigurement, treatment options are limited and generally disappointing.

[The quality of life of individuals with chronic cardiovascular diseases and diabetes mellitus]. / Qualidade de vida dos indivíduos com doenças cardiovasculares crônicas e diabetes mellitus.

The purpose of this study was to verify the chronically ill patient's quality of life. The study conceptual framework was deryged from the quality of life conceptualization according to Wenger et al. (1984). In this framework the main components of quality of life were viewed as a functional capacity, perception and symptoms. The study sample consisted of 94 chronically ill patients. Of these, 35 were in and outpatients with cardiovascular diseases (GROUP I), 29 patients were on a special rehabilitation program for cardiovascular patients (GROUP II), and 30 were diabetic patients (GROUP III). The McMaster Health Index questionnaire developed by Chambers (1984) was used for data collection. The findings revealed that patients of the GROUP II showed better quality of life than the patients of GROUP I and GROUP III in relation to the physical function, health perception, and life satisfaction.

[Structure of the dermo-epidemic junction]. / Estructura de la unión dermoepidérmica.

The basement membranes are extracellular proteic matrixes involved mainly in cell-substrate interactions. The dermo-epidermal junction (DEJ) is defined as the region formed by the dermal pole of the basal cell layer, the intercellular spaces, the lamina densa or basal lamina and the acellular fibrous components of the papillary dermis. The functions of the DEJ are to bind the epithelia to the connective tissue, and to preserve the viability of the epidermis by means of connecting it to the dermis. which is in charge of providing the necessary nutrients for its normal proliferation. The purpose of the present communication is to provide a concise review of the recent knowledges about the ultrastructure of the DEJ, with special emphasis on its antigenic components.

[Immunopathogeny of pemphigus]. / La inmunopatogenia de los pénfigos.

This article reviews recent concepts of pathogenetic mechanisms in pemphigus. We describe the pathogenic effect of the autoantibodies, the animal model for the disease and the ultrastructural alterations which follow the antibody-antigen interaction. The role of complement and protease systems in development of tissue injury are also discussed.