Dietary supplementation with whey protein improves systemic microvascular function in heart failure patients: a pilot study

Endothelial dysfunction is a well-known component of the pathophysiology of heart failure (HF), with proven prognostic value. Dietary supplementation with whey protein (WP) has been widely used to increase skeletal muscle mass, but it also has vascular effects, which are less understood. This study aimed to evaluate the effects of WP supplementation on the systemic microvascular function of HF patients. This was a blinded, randomized, placebo-controlled clinical trial that evaluated the effects of 12-week WP dietary supplementation on systemic microvascular function, in patients with HF New York Heart Association (NYHA) classes I/II. Cutaneous microvascular flow and reactivity were assessed using laser speckle contrast imaging, coupled with pharmacological local vasodilator stimuli. Fifteen patients (aged 64.5±6.2 years, 11 males) received WP supplementation and ten patients (aged 68.2±8.8 years, 8 males) received placebo (maltodextrin). The increase in endothelial-dependent microvascular vasodilation, induced by skin iontophoresis of acetylcholine, was improved after WP (P=0.03) but not placebo (P=0.37) supplementation. Moreover, endothelial-independent microvascular vasodilation induced by skin iontophoresis of sodium nitroprusside, was also enhanced after WP (P=0.04) but not placebo (P=0.42) supplementation. The results suggested that dietary supplementation with WP improved systemic microvascular function in patients with HF.

Trade-off in plant-ant interactions: seasonal variations / Custos e benefícios nas interações planta-formigas: variação sazonal

Abstract This work evaluated the effect of seasonality on ant-plant interaction in a Seasonally Dry Tropical Forests, using as an ecological model the species Ipomoea carnea subs. fistulosa (Convolvulaceae). We performed systematic collection of ants, herbivores and leaves in marked plants, evaluated the efficiency of herbivorous capture by ants, and the effects of ant presence over the pollinator behavior and plant fitness in dry and rainy seasons. The presence of ants in the plants reduced the number of herbivores (dry season: F2.27=4.7617, p=0.0166; rainy season: F2.27=5.8655, p=0.0078). However, the capture efficiency was negatively affected by the presence of myrmecophilous larvae, so that the average of ants recruited on termite leaves was 2.06 ants per termite, the average recruitment of ants on larval leaves was 22.4 larva ants. In addition, the presence of ants reduced pollinator visits and promoted fruit reduction during the dry season (ANOVA: F = 3.44; p = 0.0653). In conclusion, the association with ants can result in a balance not always favorable to the host plant, and this result actually depends on abiotic (e.g. precipitation) and biotic factors (e.g. ant species composition and abundance, influence of other trophic levels and identity of associated herbivores).

Resumo Este trabalho avaliou o efeito da sazonalidade sobre a interação planta-formiga em uma Floresta Tropical Sazonal Seca, utilizando como modelo ecológico a espécie Ipomoea carnea subs. fistulosa (Convolvulaceae). Realizamos coleta sistemática de formigas, herbívoros e folhas em plantas marcadas, avaliamos a eficiência de captura de herbívoros pelas formigas, e efeitos da presença de formigas sobre o comportamento dos polinizadores e fitness da planta nas estações seca e chuvosa. A presença de formigas nas plantas reduziu número de herbívoros (estação seca: F2.27 = 4.7617, p = 0.0166; estação chuvosa: F2.27 = 5.8655, p = 0.0078). No entanto, a eficiência de captura foi afetada negativamente pela presença de mirmecófilos larvas, enquanto a média de formigas recrutadas em folhas com cupins foi de 2,06 formigas por cupim, a média de recrutamento de formigas nas folhas com larvas foi de 22,4 formigas de larva. Além disso, a presença de formigas reduziu as visitas aos polinizadores e promoveu a redução de frutos durante a estação seca (F = 3.44, p = 0.0653). Em conclusão, a associação com formigas pode resultar em um balanço nem sempre favorável à planta hospedeira, e este resultado depende de fato de fatores abióticos (e. g. precipitação) e fatores bióticos (e.g. composição e abundância de espécies de formigas, influência de outros níveis tróficos, e identidade dos herbívoros associados).

Imunogenicidade da proteína M recombinante de Streptococcus equi subsp. equi coadministrada com um adjuvante molecular / Immunogenicity of recombinant M protein of Streptococcus equi subsp. equi co administered with a molecular adjuvant

The strangles is an infectious disease that affects horses from all ages and causes important economic losses in the equine-related business. The aim of this work was to evaluate the immunogenicity of the recombinant M protein from Streptococcus equi (rSeM) co-administered with the recombinant heat-labile enterotoxin B subunit from Escherichia coli (rLTB) in mice and horses. A total of 72 female Balb-c mice were divided into eight groups and 18 horses were divided into six groups. The animals were inoculated by intramuscular (IM) or intranasal (IN) routes with different treatments of rSeM, rLTB and/or Al(OH)3. The results obtained in both species, independent of administration routes, demonstrated that rSeM + rLTB had higher levels of specific serum immunoglobulins, however, in mucosal immunity the increase was not identified. Thus, the use of rSeM as vaccine antigen and rLTB as adjuvant can be a potential tool in the control of equine strangles.(AU)

Trade-off in plant-ant interactions: seasonal variations

Abstract This work evaluated the effect of seasonality on ant-plant interaction in a Seasonally Dry Tropical Forests, using as an ecological model the species Ipomoea carnea subs. fistulosa (Convolvulaceae). We performed systematic collection of ants, herbivores and leaves in marked plants, evaluated the efficiency of herbivorous capture by ants, and the effects of ant presence over the pollinator behavior and plant fitness in dry and rainy seasons. The presence of ants in the plants reduced the number of herbivores (dry season: F2.27=4.7617, p=0.0166; rainy season: F2.27=5.8655, p=0.0078). However, the capture efficiency was negatively affected by the presence of myrmecophilous larvae, so that the average of ants recruited on termite leaves was 2.06 ants per termite, the average recruitment of ants on larval leaves was 22.4 larva ants. In addition, the presence of ants reduced pollinator visits and promoted fruit reduction during the dry season (ANOVA: F = 3.44; p = 0.0653). In conclusion, the association with ants can result in a balance not always favorable to the host plant, and this result actually depends on abiotic (e.g. precipitation) and biotic factors (e.g. ant species composition and abundance, influence of other trophic levels and identity of associated herbivores).

Resumo Este trabalho avaliou o efeito da sazonalidade sobre a interação planta-formiga em uma Floresta Tropical Sazonal Seca, utilizando como modelo ecológico a espécie Ipomoea carnea subs. fistulosa (Convolvulaceae). Realizamos coleta sistemática de formigas, herbívoros e folhas em plantas marcadas, avaliamos a eficiência de captura de herbívoros pelas formigas, e efeitos da presença de formigas sobre o comportamento dos polinizadores e fitness da planta nas estações seca e chuvosa. A presença de formigas nas plantas reduziu número de herbívoros (estação seca: F2.27 = 4.7617, p = 0.0166; estação chuvosa: F2.27 = 5.8655, p = 0.0078). No entanto, a eficiência de captura foi afetada negativamente pela presença de mirmecófilos larvas, enquanto a média de formigas recrutadas em folhas com cupins foi de 2,06 formigas por cupim, a média de recrutamento de formigas nas folhas com larvas foi de 22,4 formigas de larva. Além disso, a presença de formigas reduziu as visitas aos polinizadores e promoveu a redução de frutos durante a estação seca (F = 3.44, p = 0.0653). Em conclusão, a associação com formigas pode resultar em um balanço nem sempre favorável à planta hospedeira, e este resultado depende de fato de fatores abióticos (e. g. precipitação) e fatores bióticos (e.g. composição e abundância de espécies de formigas, influência de outros níveis tróficos, e identidade dos herbívoros associados).

Desempenho de frangos caipiras alimentados com rações contendo diferentes níveis de energia metabolizável / Performance of broiler chickens fed diets containing different levels of metabolizable energy

Analisaram-se os efeitos da energia metabolizável (EM) e da idade de abate sobre o desempenho zootécnico, o rendimento de carcaça e de partes de frangos da linhagem Caipira Francês Exótico. Foram utilizadas 192 aves, distribuídas em delineamento inteiramente ao acaso, com três tratamentos e quatro repetições de 16 aves cada. Os tratamentos (T) foram definidos de acordo com os níveis de energia das dietas inicial (1 a 28 dias) e final (29 a 90 dias), respectivamente, em: T1 - 3.000 e 3.100kcal de EM/kg; T2 - 3.100 e 3.200kcal de EM/kg; T3 - 3.200 e 3.300kcal de EM/kg. Os abates foram realizados aos 77, 84 e 90 dias. Na fase inicial, as aves do T2 e do T3 apresentaram maior peso médio (655,75 e 713,50g), maior ganho de peso (24,28 e 26,42g/ave) e melhor conversão alimentar (1,59 e 1,77), e não diferiram entre si (P>0,05). Quando abatidas aos 77 dias de idade, as aves que consumiram a dieta com 3.100/3.200kcal de EM/kg apresentaram maior rendimento de carcaça (82,20%) e de peito (24,26%), e não diferiram dos frangos alimentados com dietas contendo 3.000/3.200kcal de EM/kg, nos quais se verificaram rendimento de carcaça de 80,72% e 23,20% de rendimento de peito. Os frangos da linhagem Caipira Francês Exótico apresentaram melhores desempenho zootécnico e rendimento de carcaça quando alimentados com dietas contendo 3.100 a 3.200kcal de EM/kg na ração e abatidos aos 77 dias de idade.

This is study aimed to analyze the effects of metabolizable energy (ME) and age at slaughter on growth performance, carcass yield and prime cuts, relative weight of the wings, back, head + neck and feet of broiler chickens from the Caipira Francês Exótico strain. A total of 192 birds were distributed in a completely randomized design with three treatments and four replicates of 16 birds per treatment. The treatments were defined according to levels of dietary energy (1 to 28 days) and late (29-90 days), respectively: T1 - 3000 and 3100kcal/kg ME, T2 - 3100 and 3200kcal/kg ME, T3 - 3200 and 3300kcal/kg ME. The slaughter was carried out at 77, 84 and 90 days. In the initial phase the birds from T2 and T3 had a higher mean weight (655.75 and 713.50g), weight gain (24.28 and 26.42g/bird) and better feed gain (1.59 and 1.77) and did not differ (P>0.05). When slaughtered at 77 days of age, birds fed the diet with 3.100/3.200kcal/kg ME had higher carcass yield (82.20%) and breast (24.26%), and did not differ from chickens fed diets containing 3.000/3.200kcal/kg ME, where a carcass yield of 80.72% and 23.20% of breast yield was found. The Caipira Francês Exótico broilers had better growth performance and carcass yield than when fed diets containing 3100-3200kcal/kg in the diet and slaughtered at 77 days old.

Do whole-body vibration exercise and resistance exercise modify concentrations of salivary cortisol and immunoglobulin A?

A single bout of resistance exercise (RE) induces hormonal and immune responses, playing an important role in a long-term adaptive process. Whole-body vibration (WBV) has also been shown to affect hormonal responses. Evidence suggests that combining WBV with RE may amplify hormonal and immune responses due to the increased neuromuscular load. Therefore, the aim of this study was to evaluate salivary cortisol (Scortisol) and salivary IgA (SIgA) concentrations following a RE session combined or not with WBV. Nine university students (22.9 ± 5.1 years, 175.8 ± 5.2 cm, and 69.2 ± 7.3 kg) performed five sets of squat exercise (70 percent one-repetition-maximum) combined (R+V30) or not (R) with WBV at 30 Hz. Saliva samples were obtained before and after exercise. Subjects also rated their effort according to the Borg CR-10 scale (RPE). Data were analyzed by a mixed model. RPE was higher after R+V30 (8.3 ± 0.7) compared to R (6.2 ± 0.7). However, Scortisol (pre: 10.6 ± 7.6 and 11.7 ± 7.6, post: 8.3 ± 6.3 and 10.2 ± 7.2 ng/mL for R and R+V30, respectively) and SIgA concentrations (pre: 98.3 ± 22.6 and 116.1 ± 51.2, post: 116.6 ± 64.7 and 143.6 ± 80.5 µg/mL for R and R+V30, respectively) were unaffected. No significant correlations were observed between Scortisol and RPE (r = 0.45, P = 0.22; r = 0.30, P = 0.42, for R and R+V30, respectively). On the basis of these data, neither protocol modified salivary cortisol or IgA, although RPE was higher after R+V30 than R.

Estudo in vitro da atividade do extrato etanólico de sementes de bacupari (Rheedia gardneriana Planch. & Triana) e das frações no crescimento de Streptococcus mutans / In vitro study of the activity of ethanol extract from bacupari (Rheedia gardneriana Planch. & Triana) seeds and its fractions on Streptococcus mutans growth

Streptococcus mutans, principal microrganismo da cavidade oral, desempenha papel preponderante na formação de placas dentárias, sendo considerado o agente etiológico primário da cárie. Rheedia gardneriana, conhecida popularmente como bacupari, é uma planta utilizada com fins medicinais para o tratamento de diversas patologias, e por apresentar atividade antimicrobiana de compostos das folhas contra bactérias Gram-positivas e Gram-negativas. O objetivo do presente trabalho foi avaliar o efeito de extrato de semente de R. gardneriana sobre a cepa S. mutans UA159. Os testes foram conduzidos com o extrato etanólico bruto e as frações obtidas com os solventes diclorometano, etanol-água, metanol e hexano, em ensaios de inibição in vitro. O extrato bruto (100 por cento) apresentou halos de inibição com diâmetro similar ao obtido com solução de digluconato de clorexidina 0,12 por cento, usada como controle. Os ensaios com a fração diclorometano exibiram atividade inibitória 35 por cento menor comparado com o controle, enquanto nenhum efeito antimicrobiano foi observado com a fração etanol-água. Contrariamente, os resultados obtidos com as frações hexânica e metanólica demonstraram claramente a atividade antimicrobiana por inibição do crescimento bacteriano. Na fração metanólica a formação de halos de inibição foi similar ao do controle. Estes dados apresentam atividade antimicrobiana de R. gardneriana contra S. mutans.

Streptococcus mutans, which is the main microorganism of the oral cavity, plays a preponderant role in dental plaque formation and is considered the primary etiologic agent regarding caries. Commonly known as "bacupari", Rheedia gardneriana is a plant used for medicinal purposes in the treatment of several pathologies; besides, its leaves have compounds that present antimicrobial activity against Gram-positive and Gram-negative bacteria. The aim of this work was to evaluate the effect of R. gardneriana seed extract on S. mutans strain UA159. The tests were carried out with crude ethanol extract and the fractions obtained with the solvents dichloromethane, ethanol-water, methanol, and hexane in in vitro inhibition assays. The crude extract (100 percent) presented inhibition halos with diameter similar to that obtained by using 0.12 percent chlorhexidine digluconate solution as control. Assays with the fraction dichloromethane showed an inhibitory activity 35 percent lower than that of the control, whereas no antimicrobial effect was observed with the ethanol-water fraction. Conversely, the results obtained with the fractions hexane and methanol clearly demonstrated antimicrobial activity by inhibiting the bacterial growth. In the methanol fraction, the formation of inhibition halos was similar to that in the control. These data present antimicrobial activity of R. gardneriana against S. mutans.

Fish oil ingestion reduces the number of aberrant crypt foci and adenoma in 1, 2-dimethylhydrazine-induced colon cancer in rats

We determined the effect of fish oil (FO) ingestion on colonic carcinogenesis in rats. Male Wistar rats received 4 subcutaneous injections (40 mg/kg body weight each) of 1,2-dimethylhydrazine (DMH) at 3-day intervals and were fed a diet containing 18 percent by weight FO (N = 10) or soybean oil (SO, N = 10) for 36 weeks. At sacrifice, the colon was removed, aberrant crypt foci were counted and the fatty acid profile was determined. Intestinal tumors were removed and classified as adenoma or carcinoma. Liver and feces were collected and analyzed for fatty acid profile. FO reduced the mean (± SEM) number of aberrant crypt foci compared to SO (113.55 ± 6.97 vs 214.60 ± 18.61; P < 0.05) and the incidence of adenoma (FO: 20 percent vs SO: 100 percent), but carcinoma occurred equally in FO and SO rats (2 animals per group). The polyunsaturated fatty acid (PUFA) profile of the colon was affected by diet (P < 0.05): total ù-3 (FO: 8.18 ± 0.97 vs SO: 1.71 ± 0.54 percent) and total ù-6 (FO: 3.83 ± 0.59 vs SO: 10.43 ± 1.28 percent). The same occurred in the liver (P < 0.05): total ù-3 (FO: 34.41 ± 2.6 vs SO: 6.46 ± 0.59 percent) and total ù-6 (FO: 8.73 ± 1.37 vs SO: 42.12 ± 2.33 percent). The PUFA profile of the feces and liver polyamine levels did not differ between groups (P > 0.05). In conclusion, our findings indicate that chronic FO ingestion protected against the DMH-induced preneoplastic colon lesions and adenoma development, but not against carcinoma in rats.

Flow rate, pH and calcium concentration of saliva of children and adolescents with type 1 diabetes mellitus

Alterations in salivary parameters may increase the caries risk in diabetic children, but, contradictory data on this issue have been reported. The aims of this study were to compare salivary parameters (flow rate, pH and calcium concentration) between healthy and type 1 diabetes mellitus (T1DM) individuals. The sample consisted of 7- to 18-year-old individuals divided into two groups: 30 subjects with T1DM (group A) and 30 healthy control subjects (group B). Fasting glucose levels were determined. Unstimulated and stimulated saliva was collected. The pH of unstimulated saliva was measured with paper strips and an electrode. Calcium concentrations in stimulated saliva were determined with a selective electrode. Group A individuals had inadequate blood glucose control (HbA1C >9%), with means ± SD unstimulated salivary flow rate of 0.15 ± 0.1 mL/min compared to 0.36 ± 0.2 mL/min for group B (P < 0.01). Stimulated salivary flow rate was similar by both groups and above 2.0 mL/min. Saliva pH was 6.0 ± 0.8 for group A and significantly different from 7.0 ± 0.6 for group B (P < 0.01). Salivary calcium was 14.7 ± 8.1 mg/L for group A and significantly higher than 9.9 ± 6.4 mg/L for group B (P < 0.01). Except for elevated calcium concentrations in saliva, salivary parameters favoring caries such as low saliva pH and unstimulated salivary flow rate were observed in T1DM individuals.

The importance of serological assays in diagnosing acute pulmonary histoplasmosis

Histoplasmosis is a systemic mycosis caused by inhalation of Histoplasma capsulatum microconidia. The disease does not normally affect immunocompetent individuals after a single, transient inhalation exposure. However, longer exposure may cause chronic or disseminated acute pulmonary infection. Herein, we report the case of a 24-year-old immunocompetent patient, who presented fever, cough and dyspnea for one month. The chest radiography revealed interstitial infiltrate and diffuse micronodules. The patient reported having had close and prolonged contact with bats. Diagnosis was confirmed by positive double immunodifusion and immunoblotting assays. She was treated with ketoconazole (400 mg) and there was complete resolution of the disease.

Long-term aerobic swimming training by rats reduces the number of aberrant crypt foci in 1, 2-dimethylhydrazine-induced colon cancer

We determined the effect of long-term aerobic swimming training regimens of different intensities on colonic carcinogenesis in rats. Male Wistar rats (11 weeks old) were given 4 subcutaneous injections (40 mg/kg body weight each) of 1,2-dimethyl-hydrazine (DMH, dissolved in 0.9 percent NaCl containing 1.5 percent EDTA, pH 6.5), at 3-day intervals and divided into three exercise groups that swam with 0 percent body weight (EG1, N = 11), 2 percent body weight (EG2, N = 11), and 4 percent body weight of load (EG3, N = 10), 20 min/day, 5 days/week for 35 weeks, and one sedentary control group (CG, N = 10). At sacrifice, the colon was removed and counted for tumors and aberrant crypt foci. Tumor size was measured and intra-abdominal fat was weighed. The mean number of aberrant crypt foci was reduced only for EG2 compared to CG (26.21 ± 2.99 vs 36.40 ± 1.53 crypts; P < 0.05). Tumor incidence was not significantly different among groups (CG: 90 percent; EG1: 72.7 percent; EG2: 90 percent; EG3: 80 percent). Swimming training did not affect either tumor multiplicity (CG: 2.30 ± 0.58; EG1: 2.09 ± 0.44; EG2: 1.27 ± 0.19; EG3: 1.50 ± 0.48 tumors) or size (CG: 1.78 ± 0.24; EG1: 1.81 ± 0.14; EG2: 1.55 ± 0.21; EG3: 2.17 ± 0.22 cm³). Intra-abdominal fat was not significantly different among groups (CG: 10.54 ± 2.73; EG1: 6.12 ± 1.15; EG2: 7.85 ± 1.24; EG3: 5.11 ± 0.74 g). Aerobic swimming training with 2 percent body weight of load protected against the DMH-induced preneoplastic colon lesions, but not against tumor development in the rat.

Influence of agitation and aeration in xanthan production by Xanthomonas campestris pv pruni strain 101 / Influencia de la agitación y la aireación en la producción de xantano por Xanthomonas campestris pv. pruni cepa 101

Production, viscosity, and chemical composition of xanthan synthesized by bacterium Xanthomonas campestris pv pruni strain 101 were evaluated in bioreactor systems. During the process, the volumetric oxygen mass transfer coefficient (kLa) and the biomass were determined and the pH was monitored. The cultures were grown in a 3 l bioreactor, with aeration and agitation varying as follows: conditions (A) 300 rpm, 3 vvm and (B) 200 rpm, 2 vvm, at 28 °C. Our results showed that gum production was dependent on kLa, with a maximum yield of 8.15 g/l at 300 rpm, 3 vvm, 54 h of fermentation, kLa 21.4/h, while biomass was not affected. All aqueous solutions of 3% (w/v) xanthans synthesized showed a pseudoplastic behavior. The highest viscosity was reached under the strongest aeration/agitation conditions. All xanthan samples contained glucose, mannose, rhamnose, and glucuronic acid as their main components. The highest agitation and aeration rates used under condition A (300 rpm and 3 vvm) favorably influenced the yield and viscosity of the xanthan produced by bacterium X. campestris pv pruni 101 at different fermentation times.

Se evaluó la producción, viscosidad y composición química del xantano sintetizado por la bacteria Xanthomonas campestris pv. pruni cepa 101 en un fermentador. Durante el proceso se controló el pH y se determinaron el coeficiente de transferencia de masa de oxígeno (kLa) y la producción de masa celular seca. Los cultivos se realizaron en un fermentador de 3 l variando la aireación y la agitación, en las siguientes condiciones: (A) 300 rpm, 3 vvm y (B) 200 rpm, 2 vvm; a 28 °C. Nuestros resultados mostraron que la producción de goma fue dependiente del kLa, con un rendimiento máximo de 8,15 g/l a 300 rpm y 3 vvm a las 54 h de fermentación, kLa de 21,4/h, mientras que la producción de biomasa no se afectó. Todas las soluciones acuosas de xantano al 3% (m/v) sintetizadas presentaron comportamiento pseudoplástico. La mayor viscosidad se alcanzó en la condición de aireación/agitación más intensa. Todas las muestras de xantano contenían glucosa, manosa, ramnosa y ácido glucurónico como constituyentes principales. La mayor tasa de agitación y aireación utilizada en la condición A (300 rpm y 3 vvm) influyó favorablemente en el rendimiento y la viscosidad del xantano producido por la bacteria X. campestris pv. pruni 101 a diferentes tiempos de fermentación.

Use of immunoblotting assay improves the sensitivity of paracoccidioidomycosis diagnosis

The purpose of this work was to evaluate two serological assays: double immunodiffusion (DI) and immunoblotting (IB) in immunodiagnosis of paracoccidioidomycosis (PCM). We evaluated by IB assay 23 sera samples from patients with clinical confirmation of PCM, all of them with negative DI results against culture filtrate from Paracoccidioides brasiliensis isolate 113. For IB, as well as for comparative DI assay, we employed soluble components of the cell wall outer surface (SCCWOS) from P. brasiliensis isolate 113 cultivated at 36°C in Fava-Neto's agar medium for 5 and 10 days. Among the 20 sera samples analyzed by DI, 13 (65 percent) were negative and 7 (35 percent) were positive against SCCWOS obtained on the 5th and 10th days. By IB assay, 95.4 percent and 100 percent of sera reacted against gp43 and gp70 present in SCCWOS from the 5th day and 95.6 percent recognized these fractions when evaluated against SCCWOS from the 10th day. Our results demonstrated that the use of an immunoenzymatic assay significantly improves the sensitivity of PCM immunodiagnosis and also suggests that at least two serological tests for antibody detection should be adopted in cases of questionable diagnosis.

Phenotype and genotype correlation of the microconversion from the CYP21A1P to the CYP21A2 gene in congenital adrenal hyperplasia

Deficiency of 21-hydroxylase is the most common form of congenital adrenal hyperplasia (CAH-21OH). We determined by allele-specific PCR the frequency of microconversion in the CYP21A2 gene in 50 Brazilian patients with the classical (salt wasting: SW and simple virilizing: SV) forms and nonclassical (NC) form of CAH-21OH and correlated genotype with phenotype. Genotypes were classified into three mutation groups (A, B, and C) based on the amount of enzymatic activity in in vitro studies using adrenal cells. In 94 unrelated alleles, we diagnosed 76 percent of the affected alleles after screening for 7 microconversions. The most frequent point mutations observed in this series were I172N (19 percent), V281L (18 percent), and IVS2,A/C>G,-12 (15 percent). In the SW form, the most frequent mutation was IVS2,A/C>G,-12 (38 percent), in the SV form it was I172N (53 percent), and in the NC form it was V281L (57.7 percent). We observed a good correlation between genotype and phenotype. Discordance between genotype and phenotype was found in one SV patient with a mild mutation in one of the alleles (R356W/V281L). However, we cannot rule out the presence of an additional mutation in these alleles. We also observed a good correlation of genotype with 17alpha-hydroxyprogesterone, testosterone, and androstenedione levels. The severity of external genitalia virilization correlated with the severity of mutation. In conclusion, the frequencies described in the present study did not differ from worldwide studies, including the Brazilian population. The few differences observed may reflect individual sample variations. This new Brazilian cohort study suggests the presence of new mutations in Brazilian patients with different forms of CAH-21OH

Polymorphism of the human immunodeficiency virus type 1 in Brazil: genetic characterization of the nef gene and implications for vaccine design

Most of the Brazilian HIV-1 samples have been characterized based on the structural genes (env, gag and pol) and no data concerning the variability of the accessory genes such as nef have been available so far. Considering the role of the nef on virus biology and the inclusion of this region in some HIV/AIDS vaccine products under testing, the purpose of this study was to document the genetic diversity of the nef gene in third-four HIV-1 Brazilian samples previously subtyped based on the env C2-V3 region. Although only few non-subtype B samples have already been analyzed so far, the cytotoxic Tlymphocyte epitopes encoded in this region were relatively conserved among the subtypes, with some amino acid signatures mainly in the subtype C samples. Considering the increasing of the non-B HIV-1 subtypes worldwide, in special the subtype C, more data should be generated concerning the genetic and antigenic variability of these subtypes, as well as the study of the impact of such polymorphism in HIV/AIDS vaccine design and testing

Parathyroid hormone secretion in chronic human endogenous hypercortisolism

Osteoporosis is a common manifestation of Cushing's syndrome, but the mechanisms responsible for this abnormality have not been defined. With the objective of analyzing parathyroid hormone (PTH) secretion in chronic hypercortisolism (CH), we evaluated 11 healthy subjects and 8 patients with CH, 6 with Cushing's disease and 2 with adrenal adenoma. These volunteers were submitted to tests of PTH stimulation through hypocalcemia (EDTA), PTH suppression through hypercalcemia (iv and oral calcium), and evaluation of bone mineral density (BMD) by DEXA. During the test of PTH stimulation, the calcium and magnesium concentrations of the normal and CH groups were similar. Patients with CH showed an increased PTH response to the hypocalcemic stimulus compared to controls. PTH values were significantly higher in the CH group at 70 (17.5 ± 3.5 vs 10.2 ± 1.3 pmol/l, P = 0.04), and 120 min (26.1 ± 5.9 vs 11.3 ± 1.9 pmol/l, P = 0.008) of EDTA infusion. The area under the curve for PTH during EDTA infusion was also significantly higher in patients with CH than in normal subjects (1867 ± 453 and 805 ± 148 pmol l-1 2 h-1, P = 0.02). During the test of PTH suppression, calcium, magnesium and PTH levels of the patients with hypercortisolism and controls were similar. BMD was decreased in patients with hypercortisolism in the spine (0.977 ± 0.052 vs 1.205 ± 0.038 g/cm² in controls, P<0.01). In conclusion, our results show that subjects with CH present decreased bone mass mainly in trabecular bone. The use of dynamic tests permitted the detection of increased PTH secretion in response to a hypocalcemic stimulus in CH patients that may probably be involved in the occurrence of osteoporosis in this state

Estudio clínico y microbiológico de la enfermedad periodontal del adulto / Clinical and microbiological study of adult periodontal disease

The aim of this study was to carry out a microbiological evaluation of sites with and without clinical evidence of moderate and severe periodontitis and their correlation with clinical parameters. A total of 52 disease sites and 10 healthy sites were selected according to clinical criteria. The following clinical indexes were measured for all the sites: plaque index, gingival index, blood on probing, depth on probing and insertion level. Samples of subgingival plaque were collected for culture and for differential counts of microbial morphotypes. In disease sites the most frequently isolated were: Prevotella intermedia/nigrescens (65), Porphyromonas gingivalis (23), Actinobacillus actinomycetemcomitans (23), Fusobacterium nucleatum (10) and Peptostreptococcus sp. (31). The aerobic gram-positive microflora was predominant in healthy sites. Significant differences were observed in microbial morphotypes between healthy and disease sites: cocci 18.71 and 78.90, motile rods 46.12 and 16.70, total spirochetes 26.48 and 2.80, respectively. The presence of motile rods, spirochetes and P. intermedia/nigrescens were the parameters with most sensitivity to suspect periodontal disease. There were significant differences in the subgingival microflora between healthy and disease sites in patients with moderate and severe periodontitis.

Salivary cortisol as a tool for physiological studies and diagnostic strategies

Salivary cortisol is an index of plasma free cortisol and is obtained by a noninvasive procedure. We have been using salivary cortisol as a tool for physiological and diagnostic studies, among them the emergence of circadian rhythm in preterm and term infants. The salivary cortisol circadian rhythm in term and premature infants was established between 8 and 12 postnatal weeks. In the preterm infants the emergence of circadian rhythm was parallel to the onset of sleep rhythm. We also studied the use of salivary cortisol for screening for Cushing's syndrome (CS) in control and obese outpatients based on circadian rhythm and the overnight 1 mg dexamethasone (DEX) suppression test. Salivary cortisol was suppressed to less than 100 ng/dl after 1 mg DEX in control and obese patients. A single salivary cortisol measurement at 23:00 h and again after 1 mg DEX above the 90th percentile of the obese group values had sensitivity and specificity of 93 and 93 percent (23:00 h), and 91 and 94 percent (after DEX), respectively. The sensitivity improved to 100 percent when we combined both parameters. We also studied 11 CS children and 21 age-matched primary obese children for whom salivary cortisol sensitivity and specificity were 100/95 percent (23:00 h), and 100/95 percent (1 mg DEX), respectively. Similar to adults, sensitivity and specificity of 100 percent were obtained by combining 23:00 h and 1 mg DEX. The measurement of salivary cortisol is a useful tool for physiological studies and for the diagnosis of CS in children and adults on an outpatient basis.

Effect of supragingival plaque control on subgingival microflora and gingivo-periodontal tissues

The aim of this study was to carry out a short term evaluation of the effect of supragingival plaque control on the subgingival microflora and gingivoperiodontal tissues in patients with moderate and severe periodontitis. A total of 92 sites from 23 patients were studied; patients ranged between 35 and 60 years of age (x 46.4 +/- 1.71). Plaque Index (PI), Gingival Index (GI), Bleeding on Probing (BP), Probing Depth (PD), and Attachment Level (AL) were measured. Subgingival microflora of the same sites was studied by dark field microscopy, and cultures in non-selective and selective media for Actinobacillus actinomycetemcomitans (Aa), Prevotella intermedia/nigrescens (Pi/n), Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn). The patients were included in a supragingival plaque control program which included oral hygiene training and assessment, and professional control of supragingival plaque and calculus. After 28 days, measurement of clinical parameters and the microbiological study were repeated. The sites were divided according to baseline PD values into: Group 1 (5 mm), Group 2 (6 mm) and Group 3 (> or = 7 mm). Results revealed a significant reduction in PI scores, GI scores, PB and PD in groups 2 and 3. No significant difference in AL was observed in any of the three groups. PI scores were found to decrease significantly in group 1, and GI scores, BP and PD tended to decrease. A significant increase in coccoid cells and a significant decrease in motile rods and spirochetes, both small and total, was observed in groups 2 and 3. Group 1 showed a significant decrease in medium and total spirochetes. At the end of the study, a significant decrease in Pi/n was found in all three groups. Significant decreases were found in Pg and Fn in Groups 3 and 2 respectively. Although it holds true that supragingival plaque control reduced a potentially pathogenic subgingival microbiota, this effect was not sufficient to be compatible with healthy gingivoperiodontal tissues.