Homeopathy on cultures of Saccharomyces cerevisiaeand impact on fermentation

Studies have shownthat homeopathy modulates the activity of both single-and multi-celled organisms;therefore, we propose a study into the action of Arnica Montanaand S. cerevisiae fungus nosode on growth "in vitro", and on the fermentation of S. cerevisiaeon brewer's wort. Methods:250 µL of medication in 30% alcohol were placed in 5 mL of Sabouraud Broth (SB) or wort, with 20 µL of fungus ata McFarland standard of 0.5 and in a dilution of 1:100. Fungal growth was evaluated via spectrophotometry at 600 nm or a cell count in a Neubauer chamber in a kinetic of 1 to 5 days' incubation at 25ºC. The production of alcohol by the fungus was evaluated using the BRIX index in the samekinetic. 1x107fungi/mL were previously incubated with medication for 5 days and, afterwards, placed in 20 mL of fresh wort, incubated at 25ºC for 7 days and evaluated for growth and sugar consumption. Resultsand Discussion: The SB results revealed that after 2days incubation with Arnica30CH, an increase in fungal growth was observed (p<0.0001), whilewith nosode 6 and 30CH there was a reduction in growth after 2 and 5 days incubation (p<0.001). The fungi incubated with Arnica30CH exhibited increased sugar consumption after 2 and5 days incubation (p<0.05), while the nosode 30CH resulted in lower sugar consumption after 2 and 3 days incubation (p<0.05). The results for fungal growth and sugar consumption with the wort were similar to those using SB.The fungalcultures previously incubated with homeopathic medication and subsequent incubation with fresh wortindicated a loss of distinction, bothin terms of fungal growth and sugar consumption. This piece of data may suggest action by the homeopathic medication only when in contact with the cells. Conclusion: The treatment of the S. cerevisiae fungus using Arnica and the S. cerevisiae nosode produced a significant modulation in fungal growth and sugar consumption.

Homeopathic medicine activity on the growth of Gram-negative bacteria

Homeopathy is a therapy that uses medications prepared with infinitesimal and dynamized dilutions. Current studies demonstrate in vitro activity of homeopathy on gram-positive bacteria such as Staphylococcus aureusand Streptococcus pyogenes. Among bacterial infections, urinary tract infection (UTI) is frequent, leads to later consequences and the main causal agent is Escherichia coli(E. coli). In other publications, it has been reported inactivity of homeopathy on E. colicultures. Due to the divergence in the literature, the objective of this study was to evaluate gram-negative bacteria growth under homeopathy treatment. Methods:The medicines Atropabelladona, Cantharis, Staphysagria,and Colibacillinumwere tested at 6CH, 12CH and 30CH inE. coliATCC 25922 and EPEC (Enteropathogenic Escherichia coli) ATCC 43887. Two hundred and fifty microliters of the medicines in alcohol 30% were incubated at 37ºC with 3 mL of Müller Hinton broth (MH), 10 µL of cultures at 0.5 Macfarland and subsequent dilution at 1/10. Bacterial growth was evaluated in a spectrophotometer at 600nm, in the periods of 6, 12,and 20 hours of incubation. Resultsand Discussion:The results showed no inhibition of bacterial growth under the studied conditions. These data corroborate with studies already published that indicate the absence of action of homeopathy on E. colicultures. Considering other studies, it can be suggested that homeopathic medicines have direct activity on the growth of Gram-positive and not Gram-negative bacteria. Evaluating the two bacterial groups, it is possible to assume that the difference in homeopathy activity could be linked to differences in the bacterial wall structure. This hypothesis should be evaluated by other tests with the same bacterialstrains. Conclusion:The homeopathic medicines tested have no direct activity on Gram-negative bacteria cultures.

The importance of optimal 25-hydroxyvitamin D levels in the glycemic control of older adults with type 2 Diabetes Mellitus: Data from the study on aging and longevity EELO

INTRODUCTION: Optimal serum levels of vitamin D are of great importance, especially in populations with comorbidities such as Diabetes Mellitus (DM). OBJECTIVE: The study evaluated the relationship between hypovitaminosis D and glycemic control in older adults with type 2 DM. METHODS: Cross-sectional and prospective study, part of the EELO project (Study on Aging and Longevity), conducted in Southern Brazil. Glycated hemoglobin (diabetes ≥6.5%) and serum levels of vitamin D (25(OH)D) were evaluated. Hypovitaminosis D was determined using cutoff points <20 and <30 ng/mL). Multivariate logistic regression was used to assess the risk of having uncontrolled DM. RESULTS: Of the 120 older adults included in the study, aged between 60 and 87 years, 74.2% were women, 66.7% used hypoglycemic medications and 75.8% exhibited uncontrolled diabetes. An inverse correlation was observed between the levels of 25(OH) D and glycated hemoglobin (rS=-0.19, p=0.037), suggesting that low levels of vitamin D are associated with poor glycemic control in diabetic individuals. The prevalence of hypovitaminosis D when using the cutoff points of <20 and <30 ng/mL were 34.2% and 75.0%, respectively. The odds ratio (OR) analysis showed that individuals with 25(OH)D<20ng/mL have almost 4 times more risk of having uncontrolled DM (OR:3.94; CI95%:1.25-12.46, p=0.02) when compared to the older adults with sufficient levels of vitamin D. CONCLUSION: The results indicate that the optimal serum levels currently recommended for 25(OH)D should preferably be 30 ng/mL or higher to contribute to better glycemic control in older adults with type 2 DM.

INTRODUÇÃO: Os níveis séricos ideais de vitamina D são de grande importância, especialmente na população com comorbidades como o Diabetes Mellitus (DM). OBJETIVO: O estudo avaliou a relação entre hipovitaminose D e controle glicêmico em idosos com DM tipo 2. MÉTODOS: Estudo transversal e prospectivo, parte do projeto EELO (Estudo sobre Envelhecimento e Longevidade), no Sul do Brasil. A hemoglobina glicada (diabetes ≥6,5%) e os níveis séricos de vitamina D (25(OH)D) foram avaliados. Hipovitaminose D foi determinada usando ponto de corte <20 e <30 ng/mL. Regressão logística multivariada foi utilizada para avaliar o risco de ter DM descompensado. RESULTADOS: Dos 120 idosos incluídos no estudo, idade entre 60 a 87 anos, 74,2% eram mulheres, 66,7% faziam uso de medicamentos hipoglicemiantes e 75,8% apresentavam diabetes descompensada. Uma correlação inversa foi observada entre os níveis de 25(OH)D e hemoglobina glicada (rS=-0,19; p=0.037), sugerindo que baixos níveis de vitamina D está associado a um pior controle glicêmico em diabéticos. A prevalência de hipovitaminose D quando se utiliza ponto de corte <20 e <30 ng/mL foi de 34,2% e 75,0%, respectivamente. A análise Odds ratio (OR) mostrou que indivíduos com 25(OH)D<20 ng/mL tem quase 4 vezes mais risco de ter DM descompensado (OR:3,94; IC95%:1,25­12,46; p=0,02) quando comparado aos idosos com níveis suficientes de vitamina D. CONCLUSÃO: Os resultados indicam que os níveis sérios ideais atualmente recomendados para 25(OH)D maior ou igual a 30 ng/ml contribuem para o melhor controle glicêmico na população idosa com DM tipo 2.

Impact of physical and functional fitness on mortality from all causes of physically independent older adults.

OBJECTIVE: To provide reference values for functional fitness tests (PFFT) and verify the capability of these tests alone and grouped into a general index (GFFI-6), to predict mortality from all causes, during seven years of follow-up of physically independent older adults. METHODS: The sample consisted of 422 older adults, evaluated at baseline using six PFFTs, as well as sociodemographic, behavioral, anthropometric, and comorbidity variables. Mortality from all causes was followed for seven subsequent years. The sample was subdivided into four groups according to sex and age. Performances in the PFFT and GFFI-6 tests were ranked into "low", "regular", and "high". RESULTS: Cox proportional regression, with the adjustment of variables, indicated that the Unipedal Balance Test (BAL), Body Agility (AGI), Sit and Stand-up (SIT-SD) tests, and GFFI-6 were able to significantly predict mortality, indicating that older adults with "low" performance have, respectively, a 2.7 (CI=1.54-4.89, p = 0.01), 4.2 (CI=2.10-8.41), 2.5 (CI=1.44-4.65, p = 0.01), and 4.7 (CI=2.10-10.81, p<0.01) times higher risk of death, compared to older adults with "high" performance. CONCLUSION: BAL, AGI, and SIT-SD tests alone and tests grouped in the GFFI-6 were strong predictors of all-cause mortality in physically independent older adults.

Hypovitaminosis D, Low Bone Mineral Density, and Diabetes Mellitus as Probable Risk Factors for Benign Paroxysmal Positional Vertigo in the Elderly

Abstract Introduction Studies have found that elderly patients with benign paroxysmal positional vertigo (BPPV) may present low levels of vitamin D (25 (OH) D), changes in bone mineral density, and diabetes mellitus (DM). Objective To investigate the possible association between BPPV, bone mineral density, hypovitaminosis D, 25 (OH) D and DM. Methods The sample consisted of 109 elderly subjects. The BPPV was verified by a standardized questionnaire and the Dix-Hallpike maneuver. Blood samples were collected for the investigation of 25 (OH) D serum levels. The bone mineral density was evaluated by means of a densitometer. Diabetes mellitus verification was performed using a self-reported questionnaire. Results Of the 109 participants, 17 had BPPV. There was a statistically significant difference between BPPV and gender (p= 0.027, phi = 0.222), with female representing 88.2% of those with BPPV. In the group with BPPV, there was a statistically significant difference for the amount of vitamin D found (p= 0.001) and for age (p= 0.001). In the elderly group with DM and BPPV, a difference was found for the standard deviation of the femur (p= 0.022) with posthoc Dunn, identifying the difference between diabetics with and without BPPV (p= 0.047). Conclusion Although no association was found (25 (OH) D levels) with BPPV in the general population of this study, it was observed that there was an association with bone mineral density in the elderly group with DM and BPPV, and, in the group with BPPV, there was an association between the amount of vitamin D and age.

Hypovitaminosis D, Low Bone Mineral Density, and Diabetes Mellitus as Probable Risk Factors for Benign Paroxysmal Positional Vertigo in the Elderly.

Introduction Studies have found that elderly patients with benign paroxysmal positional vertigo (BPPV) may present low levels of vitamin D (25 (OH) D), changes in bone mineral density, and diabetes mellitus (DM). Objective : To investigate the possible association between BPPV, bone mineral density, hypovitaminosis D, 25 (OH) D and DM. Methods The sample consisted of 109 elderly subjects. The BPPV was verified by a standardized questionnaire and the Dix-Hallpike maneuver. Blood samples were collected for the investigation of 25 (OH) D serum levels. The bone mineral density was evaluated by means of a densitometer. Diabetes mellitus verification was performed using a self-reported questionnaire. Results Of the 109 participants, 17 had BPPV. There was a statistically significant difference between BPPV and gender ( p = 0.027, phi = 0.222), with female representing 88.2% of those with BPPV. In the group with BPPV, there was a statistically significant difference for the amount of vitamin D found ( p = 0.001) and for age ( p = 0.001). In the elderly group with DM and BPPV, a difference was found for the standard deviation of the femur ( p = 0.022) with posthoc Dunn, identifying the difference between diabetics with and without BPPV ( p = 0.047). Conclusion Although no association was found (25 (OH) D levels) with BPPV in the general population of this study, it was observed that there was an association with bone mineral density in the elderly group with DM and BPPV, and, in the group with BPPV, there was an association between the amount of vitamin D and age.

Bone mineral density in independent elderly women with different physical and functional profiles and vitamin D receptor gene polymorphisms / Densidade mineral óssea de idosas independentes com diferentes perfis físico-funcionais e polimorfismos do gene do receptor da vitamina D

Abstract The aim of the study was to compare BMD among physically independent elderly women with different physical-functional profiles and vitamin D receptor gene (VDR) polymorphisms, as well as to analyze the effect of the interaction between these last two aspects on BMD. Overall, 165 elderly women had BMD assessed by bone densitometry. Handgrip and lower limb strength and functional exercise capacity (6MWT) were also assessed. VDR polymorphisms (TaqI, BsmI, ApaI and FokI) were analyzed by polymerase chain reaction. For analyses, elderly women were categorized according to their performance on physical-functional tests into low performance (LP; <25th percentile), normal performance (NP; 25th percentile ≤ NP ≤ 75th percentile) and high performance (HP;> 75th percentile). Regarding functional exercise capacity, LP group showed lower BMD compared to HP and NP groups (p=0,003). With respect to handgrip strength, there was a trend for LP group to have lower bone mineral density compared to NP group (p=0.08). No differences were observed in femur and lumbar BMD in comparisons among the different VDR genotypes (0.07≤p≤0.94); among different groups regarding lower limb strength (p=0.49) and in the interaction analysis among variables (0.17≤p≤0.77). It was concluded that physically independent elderly women with low functional exercise capacity have lower bone mineral density than those classified as normal and high performance. However, apparently, there is no effect of the interaction between VDR gene polymorphisms and physical and functional factors on BMD.

Resumo O objetivo do estudo foi comparar a DMO entre idosas fisicamente independentes com diferentes perfis físico-funcionais e polimorfismos do gene do receptor da vitamina D (VDR), assim como analisar a interação entre estes dois últimos aspectos sobre a DMO. Participaram do estudo 165 idosas, as quais tiveram a DMO avaliada por meio da densitometria óssea. Foram avaliadas também a força de preensão manual (FPM), força de membros inferiores (FMI) e a capacidade funcional de exercício (TC6min). Os polimorfismos do gene VDR (TaqI, BsmI, ApaI e FokI) foram analisados pela reação em cadeia da polimerase. Para as análises, as idosas foram categorizadas de acordo com seu desempenho físico-funcional em baixo desempenho (BD; <percentil 25), desempenho normal (DN; percentil 25 ≤ DN ≤ percentil 75) e alto desempenho (AD; > percentil 75). Para a capacidade funcional de exercício, o grupo BD apresentou menor DMO em comparação ao grupo AD e DN (p=0,003). Em relação à FPM, o grupo BD apresentou uma tendência a menor DMO quando comparado ao grupo DN (p=0,08). Não foram observadas diferenças na DMO do fêmur e da lombar na comparação entre os genótipos do gene VDR (0,07≤p≤0,94); entre os grupos em relação à FMI (p=0,24) e na análise de interação entre as variáveis (0,17≤p≤0,77). Pode-se concluir que as idosas fisicamente independentes com baixa capacidade funcional de exercício apresentam menor DMO que aquelas com desempenho normal e alto desempenho. No entanto, aparentemente não existe interação entre os polimorfismos do gene VDR e o desempenho físico-funcional sobre a DMO.

Associação entre perfil lipídico, estado nutricional e consumo alimentar em idosos atendido sem unidades de saúde, Londrina, PR / Association of lipid profile, nutritional status and dietary intake in elderly primary care health units in Londrina, PR

Introdução: as dislipidemias, a obesidade, os hábitos alimentares, o estilo de vida e as características genéticas são fatores de riscos para doenças cardiovasculares. Objetivo: investigar a associação do, perfil lipídico ao estado nutricional e ao consumo alimentar em idosos. Métodos: participaram do estudo 469 idosos. Foi avaliado o estado nutricional pelo IMC, o consumo alimentar pelo recordatório de 24 horas, o perfil lipídico por método enzimático automatizado e o uso de medicação hipolipemiante. Resultados: houve alta prevalência de excesso de peso (mulheres: 51,1%; homens: 32,5%). Somente 23,2% das mulheres e 49,4% dos homens não apresentaram dislipidemias. As mulheres apresentaram valores superiores de colesterol total, LDL-c e triglicerídeos (p < 0,05) quando comparadas aos homens, mesmo no subgrupo de pacientes medicados. A dieta caracterizou-se como hiperproteica, hiperlipídica e com baixo consumo de hortaliças e frutas. Considerações finais: os resultados reforçam a importância da promoção da alimentação saudável no âmbito da atenção básica, independente do uso de medicação hipolipemiante, pois as alterações no perfil lipídico e no estado nutricional persistem, indicando risco aumentado para doenças cardiovasculares. (AU)

Introduction: dyslipidemia, obesity, unbalanced diet, life style and genetic background are risk factors for cardiovascular diseases. Objective: this study aimed to evaluate the association of lipid profi le, nutritional status and dietary intake in elderly. Methods: the studied population was composed by 469 individuals. Nutritional status was assessed by BMI, dietary intake by 24-Hour Dietary Recall, the lipid profi le by enzymatic method and use of antilipidaemic medication. Results: a high prevalence of overweight was observed (women: 51.1%; men: 32.5%). Only 23.2% of women and 49.4% of men did not have dyslipidemia. The women showed higher values of total cholesterol, LDL-c and triglycerides compared to men, even in patients using antilipidaemic medication (p < 0.05). Diet was hyperlipidemic, hyperproteic and with low vegetables consumption. Final remarks: these results reinforce the signifi cance of healthy diet in primary attention despite the use of antilipidemic medication, since changes in the lipid profi le and nutritional status are sustained indicating an increased risk for cardiovascular diseases. (AU)

Associação entre perfil lipídico, estado nutricional e consumo alimentar em idosos atendido sem unidades de saúde, Londrina, PR / Association of lipid profile, nutritional status and dietary intake in elderly primary care health units in Londrina, PR

Introdução: as dislipidemias, a obesidade, os hábitos alimentares, o estilo de vida e as características genéticas são fatores de riscos para doenças cardiovasculares. Objetivo: investigar a associação do, perfil lipídico ao estado nutricional e ao consumo alimentar em idosos. Métodos: participaram do estudo 469 idosos. Foi avaliado o estado nutricional pelo IMC, o consumo alimentar pelo recordatório de 24 horas, o perfil lipídico por método enzimático automatizado e o uso de medicação hipolipemiante. Resultados: houve alta prevalência de excesso de peso (mulheres: 51,1%; homens: 32,5%). Somente 23,2% das mulheres e 49,4% dos homens não apresentaram dislipidemias. As mulheres apresentaram valores superiores de colesterol total, LDL-c e triglicerídeos (p < 0,05) quando comparadas aos homens, mesmo no subgrupo de pacientes medicados. A dieta caracterizou-se como hiperproteica, hiperlipídica e com baixo consumo de hortaliças e frutas. Considerações finais: os resultados reforçam a importância da promoção da alimentação saudável no âmbito da atenção básica, independente do uso de medicação hipolipemiante, pois as alterações no perfil lipídico e no estado nutricional persistem, indicando risco aumentado para doenças cardiovasculares.

Introduction: dyslipidemia, obesity, unbalanced diet, life style and genetic background are risk factors for cardiovascular diseases. Objective: this study aimed to evaluate the association of lipid profi le, nutritional status and dietary intake in elderly. Methods: the studied population was composed by 469 individuals. Nutritional status was assessed by BMI, dietary intake by 24-Hour Dietary Recall, the lipid profi le by enzymatic method and use of antilipidaemic medication. Results: a high prevalence of overweight was observed (women: 51.1%; men: 32.5%). Only 23.2% of women and 49.4% of men did not have dyslipidemia. The women showed higher values of total cholesterol, LDL-c and triglycerides compared to men, even in patients using antilipidaemic medication (p < 0.05). Diet was hyperlipidemic, hyperproteic and with low vegetables consumption. Final remarks: these results reinforce the signifi cance of healthy diet in primary attention despite the use of antilipidemic medication, since changes in the lipid profi le and nutritional status are sustained indicating an increased risk for cardiovascular diseases.

Prevalence of risk factors for the occurrence of strokes in the elderly / Prevalência de fatores de risco para a ocorrência de acidente vascular encefálico em idosos

Introduction Stroke (CVA) is a multifactorial disease, where the combination of risk factors may be associated with and contribute to its development. Objectives The objective of this study was to evaluate the prevalence of risk factors for stroke in the physically independent elderly in the city of Londrina, Brazil. Materials and methods This study was composed of the elderly individuals participating in the EELO project (Age and Ageing Study, Londrina, State of Paraná, Brazil). In order to identify the risk factors for stroke, one used a structured questionnaire with information about socio-demographic and anthropometric data as well as lifestyle variables, such as physical inactivity and smoking, presence of comorbidities, and laboratory tests for diagnosis of diabetes and dyslipidemia. Results The study included 454 elderly with mean age of 69.7 years. There was a significant relationship between the number of risk factors and gender (p = 0.01), with higher prevalence among women. In separate analysis, the elderly between 60 and 69 years had a higher incidence of obesity (p = 0.03) and dyslipidemia (p = 0.04). Regarding gender, obesity (p = 0.01), smoking (p = 0.0001), vascular disease (p = 0.0001) and heart disease in the family (p = 0.01) higher incidence was shown in females, according toChi Square’s test. Conclusions It is concluded that elderly people aged less than 70 years and older women were those with the highest number of risk factors for CVA. Therefore, it may be suggested the development of primary care programs in order to promote information on the prevention of these risk factors and thus reduce the occurrence of stroke.

Introdução O Acidente Vascular Encefálico (AVE) é uma doença multifatorial, de modo que a combinação de fatores de risco pode contribuir para o seu desenvolvimento. Objetivo O objetivo deste estudo foi avaliar a prevalência dos fatores de risco para o AVE na população idosa fisicamente independente do município de Londrina. Materiais e métodos O estudo foi composto por idosos integrantes do projeto EELO (Estudo sobre o Envelhecimento e Longevidade – Londrina (PR), Brasil). Para identificação dos fatores de risco do AVE, utilizou-se um questionário estruturado com informações sobre dados sociodemográficos, dados antropométricos, variáveis de estilo de vida como sedentarismo e hábito de fumar, análise da presença de comorbidades e avaliação laboratorial para o diagnóstico de diabetes e dislipidemias. Resultados Participaram deste estudo 454 idosos com idade média de 69,7 anos. Observou-se relação significativa entre o número de fatores de risco e o gênero (p = 0,01), com maior prevalência entre as mulheres. Em análise isolada, idosos entre 60 e 69 anos apresentaram maior incidência de obesidade (p = 0,03) e dislipidemias (p = 0,04). Em relação ao gênero, a obesidade (p = 0,01), tabagismo (p = 0,0001), doença vascular (p = 0,0001) e doença cardíaca na família (p = 0,01) apresentaram maior incidência no sexo feminino, segundo teste Qui Quadrado. Conclusões Conclui-se que os idosos com idade menor que 70 anos e mulheres idosas foram os que apresentaram maior número de fatores de risco para o AVE. Desta forma sugere-se a elaboração de programas de atenção primária com o objetivo de promover informações sobre a prevenção destes fatores de risco e a redução da ocorrência do acidente vascular encefálico.

High molecular mass fraction in clinical isolates of Paracoccidioides brasiliensis.

INTRODUCTION: Different serum levels of the IgG/IgE for Paracoccidioides brasiliensis high mass molecular (hMM) fraction (~366 kDa) in the acute and chronic forms of the disease have been reported. Considering the nonexistence of hMM fraction investigation involving clinical isolates of P. brasiliensis, the present study aimed to investigate the presence of the hMM fraction (~366 kDa) in cell free antigens (CFA) from P. brasiliensis clinical isolates. METHODS: CFA from 10 clinical isolates and a reference strain (Pb18) were submitted to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by gel image capturing and densitometer analysis. Additionally, CFA from 20 isolates and Pb18 were analyzed by capture ELISA (cELISA) using polyclonal (polAb) or monoclonal (mAb) antibodies to the hMM fraction. RESULTS: The presence of the hMM component was observed in CFA of all samples analyzed by SDS-PAGE/densitometry and by cELISA. In addition, Pearson's correlation test demonstrated stronger coefficients between hMM fraction levels using pAb and mAb (R = 0.853) in cELISA. CONCLUSIONS: The soluble hMM fraction was present in all the P. brasiliensis clinical isolates analyzed and the reference strain Pb18, which could be used as a source of this antigen. The work also introduces for first time, the cELISA method for P. brasiliensis hMM fraction detection. Analysis also suggests that detection is viable using polAb or mAb and this methodology may be useful for future investigation of the soluble hMM fraction (~366 kDa) in sera from PCM patients.

High molecular mass fraction in clinical isolates of Paracoccidioides brasiliensis / Fração de alta massa molecular em isolados clínicos de Paracoccidioides brasiliensis

INTRODUCTION: Different serum levels of the IgG/IgE for Paracoccidioides brasiliensis high mass molecular (hMM) fraction (~366kDa) in the acute and chronic forms of the disease have been reported. Considering the nonexistence of hMM fraction investigation involving clinical isolates of P. brasiliensis, the present study aimed to investigate the presence of the hMM fraction (~366kDa) in cell free antigens (CFA) from P. brasiliensis clinical isolates. METHODS: CFA from 10 clinical isolates and a reference strain (Pb18) were submitted to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by gel image capturing and densitometer analysis. Additionally, CFA from 20 isolates and Pb18 were analyzed by capture ELISA (cELISA) using polyclonal (polAb) or monoclonal (mAb) antibodies to the hMM fraction. RESULTS: The presence of the hMM component was observed in CFA of all samples analyzed by SDS-PAGE/densitometry and by cELISA. In addition, Pearson's correlation test demonstrated stronger coefficients between hMM fraction levels using pAb and mAb (R = 0.853) in cELISA. CONCLUSIONS: The soluble hMM fraction was present in all the P. brasiliensis clinical isolates analyzed and the reference strain Pb18, which could be used as a source of this antigen. The work also introduces for first time, the cELISA method for P. brasiliensis hMM fraction detection. Analysis also suggests that detection is viable using polAb or mAb and this methodology may be useful for future investigation of the soluble hMM fraction (~366kDa) in sera from PCM patients.

INTRODUÇÃO: Diferentes níveis sorológicos de IgG/IgE contra a fração de alta massa molecular (hMM) (~366kDa) de Paracoccidioides brasiliensis têm sido encontrados na PCM aguda e crônica. Considerando a inexistência de investigação sobre esta fração em isolados clínicos de P. brasiliensis, o objetivo deste estudo foi investigar a presença da fração hMM (~366kDa) no preparado livre de células (CFA) de P. brasiliensis obtidos de isolados clínicos. MÉTODOS: CFA de 10 isolados e de cepa de referência (Pb18) foram submetidas à eletroforese em gel de SDS-poliacrilamida (SDS-PAGE) seguida de captura de imagem e análise por densitometria. Adicionalmente, CFA de 20 isolados e de Pb18 foram analisados por ELISA captura (cELISA) utilizando anticorpos policlonal (polAb) ou monoclonal (mAb) para fração hMM. RESULTADOS: A presença do componente de hMM foi observada em todas as amostras analisadas por SDS-PAGE/densitometria e por cELISA. Adicionalmente, o teste de correlação de Pearson demonstrou forte relação entre os níveis de fração hMM usando pAb e mAb (R = 0.853) no cELISA. CONCLUSÕES: Conclui-se que a fração hMM está presente em todos os isolados clínicos de P. brasiliensis analisados e no isolado referencial, sugerindo a possibilidade dos mesmos serem utilizados como fonte desta fração antigênica. Este trabalho também introduz pela primeira vez o método de cELISA para detecção da fração hMM de P. brasiliensis, sugerindo que detecção utilizando anticorpos polAb ou mAb é viável e essa metodologia poderá ser útil para investigação futura desta fração solúvel (~366kDa) em soros de pacientes com PCM.

Serum proteins and fractions, HDL-cholesterol and total IgG and IgE levels in cases of acute and chronic paracoccidioidomycosis.

This study evaluated serum protein fractions, HDL-cholesterol, total immunoglobulin G and total immunoglobulin E levels in patients with acute and chronic paracoccidioidomycosis, by means of electrophoresis, enzymatic reaction and immunoenzymatic assay. The results demonstrated elevated levels of total immunoglobulin G, total immunoglobulin E, alpha-2 and gamma-globulins, which were more evident in acute than in chronic PCM, but no increase in HDL-cholesterol levels. There was a correlation between the levels of total immunoglobulin E and gamma-globulins and the alpha-2 and beta-globulin fractions in the acute form and between beta and gamma-globulins in both the acute and the chronic form. In conclusion, changes in total immunoglobulin G and immunoglobulin E levels and in the electrophoretic profile may be important markers for the prognosis and therapeutic follow-up of PCM cases, especially because protein electrophoresis is a simple laboratory test that can be applied when specific PCM serological tests are not available. In addition, levels of the gamma-globulin fraction greater than 2.0 g/dl may suggest that the patient is developing a more severe form of PCM.

Serum proteins and fractions, HDL-cholesterol and total IgG and IgE levels in cases of acute and chronic paracoccidioidomycosis / Proteínas séricas e frações, HDL-colesterol e níveis de IgG e IgE totais na paracoccidioidomicose aguda e crônica

This study evaluated serum protein fractions, HDL-cholesterol, total immunoglobulin G and total immunoglobulin E levels in patients with acute and chronic paracoccidioidomycosis, by means of electrophoresis, enzymatic reaction and immunoenzymatic assay. The results demonstrated elevated levels of total immunoglobulin G, total immunoglobulin E, alpha-2 and gamma-globulins, which were more evident in acute than in chronic PCM, but no increase in HDL-cholesterol levels. There was a correlation between the levels of total immunoglobulin E and gamma-globulins and the alpha-2 and beta-globulin fractions in the acute form and between beta and gamma-globulins in both the acute and the chronic form. In conclusion, changes in total immunoglobulin G and immunoglobulin E levels and in the electrophoretic profile may be important markers for the prognosis and therapeutic follow-up of PCM cases, especially because protein electrophoresis is a simple laboratory test that can be applied when specific PCM serological tests are not available. In addition, levels of the gamma-globulin fraction greater than 2.0g/dl may suggest that the patient is developing a more severe form of PCM.

Este trabalho avaliou as frações de proteínas séricas, HDL-colesterol e imunoglobulina G e imunoglobulina E totais em pacientes com paracoccidioidomicose aguda e crônica por eletroforese, reação enzimática e ensaio imunoenzimático. Os resultados demonstraram aumento dos níveis de imunoglobulina G e imunoglobulina E totais, alfa-2 e gama-globulinas, mais evidente na forma aguda que na forma crônica, e não nos níveis de HDL-colesterol. Houve correlação entre níveis de imunoglobulina E total e gama-globulinas e fração alfa-2 e beta-globulinas na forma aguda e entre beta e gama-globulinas nas formas aguda e crônica. Concluindo, alterações nos níveis de imunoglobulina G e imunoglobulina E totais e no perfil eletroforético podem ser importantes marcadores para prognóstico e acompanhamento terapêutico da PCM, especialmente por ser a eletroforese de proteínas um exame laboratorial simples que pode ser empregado em situações onde a sorologia específica para PCM não está disponível. Adicionalmente, níveis da fração gama-globulinas acima de 2,0g/dL podem sugerir que o paciente esteja desenvolvendo uma forma mais grave de PCM.

Estudo comparativo dos diagnósticos de vaginose bacteriana pelas técnicas de Papanicolaou e Gram / Comparative study for the diagnosis of bacterial vaginosis by Papanicolaou and Gram staining techniques

Este trabalho teve como objetivo determinar a habilidade do exame citopatológico de Papanicolaou para diagnosticar a vaginose bacteriana (VB), usando a coloração de Gram (gradiente de Nugent) como método diagnóstico padrão. Em estudo prospectivo foram analisadas amostras cérvico-vaginais de 223 mulheres não gestantes, com idades entre 14 e 82 anos. Na bacterioscopia ao Gram, a VB foi definida de acordo com os critérios propostos por Nugent et al. (1991), e, na técnica de Papanicolaou, a afecção foi caracterizada pela presença de microbiota mista com predomínio de cocobacilos, presença de clue cells e ausência ou diminuição da microbiotabacilar. A leitura dos esfregaços foi realizada por dois examinadores individualmente e de forma independente. O grau de concordância entre os resultados obtidos pelos dois métodos foi definido pelo coeficiente kappa. Comparado ao Gram, a sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo do método de Papanicolaou para diagnóstico de VB foram, respectivamente, 71%, 93%, 78% e 91%. O cálculo do coeficiente kappa demonstrou boa concordância entre os dois procedimentos (k=0,7). Os resultados obtidos sugerem que a análise dos esfregaços cérvico-vaginais corados pela técnica de Papanicolaou pode representar uma alternativa prática e útil para o diagnóstico de VB.

Disúria e polaciúria: sintomas realmente sugestivos de infecção do trato urinário? / Dysuria and polaciuria: really a suggestive symptom of urinary tract infection?

Infecção do trato urinário (ITU) é considerada uma das infecções mais comuns na população, sendo responsável por significativa morbidade no sexo feminino. Na rotina dos consultórios médicos ambulatoriais, o tratamento das ITU não complicadas em mulheres adultas é iniciado com base na anamnese, já que a maioria das ITU é acompanhada por sintomas como disúria e polaciúria. O presente estudo visou determinar a correlação entre os sintomas clássicos de ITU e o diagnóstico efetivo desta patologia, caracterizado pela urocultura positiva, atravé de um inquérito epidemiológico realizado com mulheres com idade entre 15 a 55 anos. Das 191 mulheres avaliadas, 56 (29%) apresentaram sintomas sugestivos de ITU. Das 16 mulheres que apresentaram disúria, 2 (12,5%)tiveram o diagnóstico de ITU confirmnado. A presença de disúria e polaciúria ocorreu em 27 mulheres sendo que 5 (18,5%) destas apresentavam realmente ITU. Os coeficientes de correlação entre disúria, polaciúria e a soma destes sintoma com o resultado da cultura foram inferiores a 0,5, demonstrando que não ocorreu correlação significativa entre estas variáveis. Este estudo sugere que a anamnese não deve ser utilizada como critério único para diagnóstico das ITU e que este deve ser confirmado sempre que possível com exames laboratoriais.