SHIFT IN HUMAN ROTAVIRUS DISTRIBUTION IN BELO HORIZONTE, BRAZIL DETECTED BY RIBONUCLEIC ACID ELECTROPHORESIS

Rotavirus has been considered the main agent of infectious diarrhea especially among younger children. We addressed the prevalence of rotavirus-associated diarrhea and the diversity of circulating electropherotypes by immunochromatography and RNA electrophoresis. Stool samples were taken from 391 children (267 with diarrhea) from the lower socioeconomic stratum who sought treatment in the Hospital Infantil João Paulo II/Belo Horizonte, during 2005 and 2006. Rotavirus was detected in 79/20.2% of subjects, 64/24.0% with diarrhea and 15/12.1% with no diarrhea. The virus was strongly associated with diarrhea (p = 0.003). A total of 76/19.4% and 69/17.6% rotavirus-positive children were identified by immunochromatography and electrophoresis, respectively. Rotavirus-associated diarrhea was more frequently detected in dry months (p < 0.001) and almost exclusively in children aged up to three years. Long profile strains prevailed (54/78.3%) but a shift toward short electropherotype was identified. Despite the decrease seen in 2006, rotavirus infection is still very common in our area. Although viral RNA electrophoresis is useful as a typing method, it should not be used exclusively in the diagnosis of rotavirus infection. We confirmed a shift from long to short profile strains, as already described for other South American countries.

Comparação entre os métodos de Ziehl-Neelsen modificado e Acid-Fast-Trichrome para a pesquisa fecal de Cryptosporidium parvum e Isospora belli

Devido a crescente importância dos coccídios intestinais (Cryptosporidium, Isospora e Cyclospora) como parasitos oportunistas, é fundamental para os laboratórios diferenciar morfologicamente estes protozoários; a técnica de Ziehl-Neelsen modificada (ZNm) é amplamente utilizada para este fim; recentemente, foi proposto um novo procedimento, a coloração combinada do ácido tricrômico (Acid-Fast-Trichrome - AFT). O objetivo do presente estudo foi comparar os processos AFT e ZNm para a detecção destes coccídios em amostras fecais de pacientes portadores do vírus VIH. Foram selecionados dois grupos de indivíduos, para inclusão no estudo, segundo a presença (n=60) ou ausência de diarréia (n=60). As amostras de fezes foram coletadas em solução de formalina 10% e os esfregaços fecais preparados i) diretamente das fezes e ii) após concentração prévia a 500xg (10 minutos), foram submetidos aos diferentes processos de coloração. Considerando-se a positividade por técnica (AFT e ZNm), verificou-se a superioridade do procedimento de ZNm (n=19; 100% dos casos positivos) sobre o de AFT (n=8; 42,1%). Ambos possibilitaram a identificação dos 101 casos verdadeiramente negativos. Coccidiose intestinal foi mais frequente entre os pacientes que apresentaram diarréia (26,6%) em comparação à positividade observada entre os indíviduos assintomáticos (5%) sendo que C. cayetanensis não foi detectada em ambos os grupos. Foi de nosso interesse avaliar a aplicabilidade da técnica AFT para a coloração deste protozoário. Devido à sensibilidade e especificidade obtida neste estudo (100%), conclui-se que o método de ZNm continua sendo o mais indicado para o diagnóstico da criptosporidiose e isosporose, principalmente quando associado ao procedimento de centrífugo-concentração (500xg, 10 minutos). Embora a coloração AFT tenha baixo custo, faz-se necessário o seu aperfeiçoamento pois este procedimento permite o diagnóstico simultâneo dos coccídios intestinais (C. parvum, I. belli e C. cayetanensis) e dos microsporídios.

In vitro differential activity of phospholipases and acid proteinases of clinical isolates of Candida

INTRODUCTION: Candida yeasts are commensals; however, if the balance of normal flora is disrupted or the immune defenses are compromised, Candida species can cause disease manifestations. Several attributes contribute to the virulence and pathogenicity of Candida, including the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate the in vitro activity of phospholipases and acid proteinases in clinical isolates of Candida spp. METHODS: Eighty-two isolates from hospitalized patients collected from various sites of origin were analyzed. Phospholipase production was performed in egg yolk medium and the production of proteinase was verified in a medium containing bovine serum albumin. The study was performed in triplicate. RESULTS: Fifty-six (68.3%) of isolates tested were phospholipase positive and 16 (44.4%) were positive for proteinase activity. C. tropicalis was the species with the highest number of positive isolates for phospholipase (91.7%). Statistically significant differences were observed in relation to production of phospholipases among species (p<0,0001) and among the strains from different sites of origin (p=0.014). Regarding the production of acid protease, the isolates of C. parapsilosis tested presented a larger number of producers (69.2%). Among the species analyzed, the percentage of protease producing isolates did not differ statistically (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSIONS: The majority of C. non-albicans and all C. albicans isolates were great producers of hydrolytic enzymes and, consequently, might be able to cause infection under favorable conditions.

A first case of protease codon 35 amino acid insertion in a HIV-1 subtype B sequence detected in the Bauru region, state of São Paulo, Brazil: case report

Amino acid insertions in the protease have rarely been described in HIVinfected patients. One of these insertions has recently been described in codon 35, although its impact on resistance remains unknown. This study presents a case of an HIV variant with an insertion in codon 35 of the protease, described for the first time in Bauru, State of Sao Paulo, Brazil, circulating in a 38-year-old caucasian male with asymptomatic HIV infection since 1997. The variant isolated showed a codon 35 insertion of two amino acids in the protease: a threonine and an aspartic acid, resulting in the amino acid sequence E35E_TD.

In vitro antifungal activity of fatty acid methyl esters of the seeds of Annona cornifolia A.St.-Hil. (Annonaceae) against pathogenic fungus Paracoccidioides brasiliensis

INTRODUCTION: Fatty acids are abundant in vegetable oils. They are known to have antibacterial and antifungal properties. METHODS: Antifungal susceptibility was evaluated by broth microdilution assay following CLSI (formerly the NCCLS) guidelines against 16 fungal strains of clinical interest. RESULTS: In this work, fatty acid methyl esters (FAME) was able to inhibit 12 clinical strains of the pathogenic fungus Paracoccidioides brasiliensis and were also active in the bioautographic assay against Cladosporium sphaerospermum. CONCLUSIONS: FAME was a more potent antifungal than trimethoprim-sulfamethoxazole against P. brasiliensis under the experimental conditions tested.

Evaluation of bacterial growth inhibition by mercaptopropionic acid in metallo-β-lactamase detection on multidrug-resistant Acinetobacter baumannii

INTRODUCTION: Metallo-β-lactamase (MBL) has been reported all over the world. METHODS: The inhibitory effect of mercaptopropionic acid (MPA) on bacterial growth was evaluated by comparison between disk diffusion and broth dilution methodology with determination of the minimum inhibitory concentration (MIC) for multidrug-resistant Acinetobacter baumanni strains. RESULTS: MPA significantly inhibited growth of the strains. CONCLUSIONS: The use of MPA can affect the results in phenotypic methods of MBL detection.

Larvicidal activity of crude extracts from Larrea cuneifolia (Zygophyllaceae) and of its metabolite nordihydroguaiaretic acid against the vector Culex quinquefasciatus (Diptera: Culicidae)

INTRODUCTION: The aim of the present study was to analyze the larvicidal activity of different crude extracts of Larrea cuneifolia and its most abundant lignan, nordihydroguaiaretic acid (NDGA), against Culex quinquefasciatus. METHODS: Chloroform, methanol, and aqueous extracts from L. cuneifolia and NDGA were tested against larvae of Cx. quinquefasciatus under laboratory conditions. RESULTS: The chloroform extract showed the highest larvicidal effect, with an estimated LC50 of 0.062 mg/ml. NDGA also demonstrated significant larvicidal activity with an estimated LC50 of 0.092 mg/ml. CONCLUSIONS: These results indicate that the chloroform extract of L. cuneifolia and NDGA are promising insecticides of botanical origin that could be useful for controlling Cx. quinquefasciatus.

Evaluation of three enzyme immunoassays and a nucleic acid amplification test for the diagnosis of Clostridium difficile-associated diarrhea at a university hospital in Brazil

Introduction Despite the known importance of Clostridium difficile as a nosocomial pathogen, few studies regarding Clostridium difficile infection (CDI) in Brazil have been conducted. To date, the diagnostic tests that are available on the Brazilian market for the diagnosis of CDI have not been evaluated. The aim of this study was to compare the performances of four commercial methods for the diagnosis of CDI in patients from a university hospital in Brazil. Methods Three enzyme immunoassays (EIAs) and one nucleic acid amplification test (NAAT) were evaluated against a cytotoxicity assay (CTA) and toxigenic culture (TC). Stool samples from 92 patients with suspected CDI were used in this study. Results Twenty-five (27.2%) of 92 samples were positive according to the CTA, and 23 (25%) were positive according to the TC. All EIAs and the NAAT test demonstrated sensitivities between 59 and 68% and specificities greater than 91%. Conclusions All four methods exhibited low sensitivities for the diagnosis of CDI, which could lead to a large number of false-negative results, an increased risk of cross-infection to other patients, and overtreatment with empirical antibiotics.

Ursodeoxycholic acid does not interfere with in vivo Helicobacter pylori colonization

A low frequency of Helicobacter pylori in the gastric mucosa of patients with alkaline gastritis has been reported. At the same time, it can be noted that the growth of bacteria can be inhibited by bile acids. We studied 40 patients with chronic gastritis related to Helicobacter pylori in order to determine the effect of ursodeoxycholic acid on this infection. Diagnoses of the infection and the inflammatory process were obtained by histologic study of gastric biopsies collected during endoscopy. Two groups were studied: group I received ursodeoxycholic acid - 300 mg/day, and group II received the placebo, twice a day, both for 28 days. The colonization by Helicobacter pylori and the intensity of the mononuclear and polymorphonuclear inflammatory infiltrate were determined before (time 1) and after (time 2) treatment. Ursodeoxycholic acid had no effect on the Helicobacter pylori infection. A significant reduction in the intensity of the mononuclear inflammatory infiltrate of the gastric antrum mucosa was observed in patients from group I, when we compared not only times 1 and 2 but also groups I and II. However, this was not the case with the body mucosa. We concluded that ursodeoxycholic acid had no action on the colonization by Helicobacter pylori or on the polymorphonuclear inflammatory infiltrate, but it caused a significant reduction in the intensity of the mononuclear inflammatory infiltrate of the gastric antrum.

The effect of intravenous zoledronic acid on glucocorticoid-induced multiple vertebral fractures in juvenile systemic lupus erythematosus

Glucocorticoids are widely used in the treatment of lupus patients, and adverse effects, which include osteoporosis and associated fractures, are frequent. Treatment of osteoporosis of young patients should be effective and not harmful to bone growth and remodeling. Bisphosphonates are drugs that decrease the incidence of bone fractures, but their use in juvenile patients is still controversial because of their possible side effects on the growing skeleton. However, recently published studies showed that linear growth continued normally after treatment with these drugs, and there was no excessive suppression of bone remodeling or mineralization defects. Zoledronic acid is a new intravenous bisphosphonate that has been approved by the US FDA for use with hypercalcemia of malignancies and might be an effective treatment for postmenopausal osteoporosis. The authors report a case of a young girl with systemic lupus who developed multiple vertebral collapses due to glucocorticoid therapy, and zoledronic acid was used producing significant clinical and densitometric improvement.

Seleção de genótipos de pimentão resistentes à Xanthomonas campestris pv. vesicatoria (Doidge) Dye. sob condições naturais de infecção

Devido à ocorrência de epidemias severas de pústula bacteriana ou mancha bacteriana no pimentão, causada pela bactéria Xanthomonas campestris pv. vesicatoria (Doidge) Dye., o cultivo do pimentão na várzea do Rio Solimões, próximo à Manaus, encontra-se em decadência. O INPA, desde 1976, desenvolve um Programa de Melhoramento Genético do Pimentão visando incorporar resistência ao patógeno. Neste trabalho são relatados os resultados obtidos em três ensaios, nas áreas de terra firme e várzea do Estado do Amazonas, envolvendo progênies F13 e F14 do cruzamento interespecífico entre Capsicum annuum e C. chinense, denominado HP-12, em cujas progênies vêm sendo realizadas seleções genealógicas visando obter variedades resistentes ao patógeno X. campestris pv. vesicatoria e alta capacidade produtiva, sob condição de cultivo em ambientes quentes e úmidos. Quando a população de hospedeiros foi constituída por indivíduos resistentes e suscetíveis, a curva de progresso da doença adaptou-se melhor ao modelo monomolecular, onde níveis mais elevados de resistência, conferidos por um genótipo, foram devidos à sua capacidade de restringir a velocidade do progresso da doença. Nos três ensaios, as progênies selecionadas pelo Programa apresentaram maior resistência e capacidade produtiva, quando comparadas à testemunha suscetível (Cascadura Ikeda), em condições de ocorrência da doença e verificou-se que a capacidade de produção de frutos está relacionada aos níveis de resistência do hospedeiro ao patógeno. Por outro lado, levando-se em conta os caracteres de resistência e capacidade produtiva das progênies inferiu-se que a espécie C. chinense é um recurso genético importante como fonte de resistência a X. campestris pv. vesicatoria nos programas de melhoramento do pimentão.

Fatty Acid and Cholesterol Concentrations in Usually Consumed Fish in Brazil

Background: Several studies have demonstrated clinical benefits of fish consumption for the cardiovascular system. These effects are attributed to the increased amounts of polyunsaturated fatty acids in these foods. However, the concentrations of fatty acids may vary according to region. Objective: The goal of this study was to determine the amount of,cholesterol and fatty acids in 10 Brazilian fishes and in a non-native farmed salmon usually consumed in Brazil. Methods: The concentrations of cholesterol and fatty acids, especially omega-3, were determined in grilled fishes. Each fish sample was divided in 3 sub-samples (chops) and each one was extracted from the fish to minimize possible differences in muscle and fat contents. Results: The largest cholesterol amount was found in white grouper (107.6 mg/100 g of fish) and the smallest in badejo (70 mg/100 g). Omega-3 amount varied from 0.01 g/100 g in badejo to 0.900 g/100 g in weakfish. Saturated fat varied from 0.687 g/100 g in seabass to 4.530 g/100 g in filhote. The salmon had the greatest concentration of polyunsaturated fats (3.29 g/100 g) and the highest content of monounsaturated was found in pescadinha (5.98 g/100 g). Whiting and boyfriend had the best omega-6/omega 3 ratios respectively 2.22 and 1.19, however these species showed very little amounts of omega-3. Conclusion: All studied Brazilian fishes and imported salmon have low amounts of saturated fat and most of them also have low amounts of omega-3.

Effects of 2-chloroetylphosphonic acid on ripening of mango

Mature fruits of mango 'Paheri' were treated immediately after harvest with ethefon at 0 - 250 - 500 - 1.000 and 2.000 ppm. Fruit ripening was accelerated by all treatments , the time to maturity being reduced from 48 to 72 hours, when compared with controls. Maturation was evaluated, by external colour of fruits, soluble solids and acid contents.

Sobre o preparo dos parasitas da malária em distensões espessas, especialmente em amostras supersecas

I) the A. presents a method developed for the preparation of thick blood films, specially old desiccated smears. The observations are based on the experience of more than 53000 blood samples collected in the laboratory of the “Serviço de Malaria do Nordeste” as well as in the research department of the “Serviço de Malaria da Baixada Fluminenese”. II) As in introductory matter, he emphasizes the value of the obstacles presented by overdrying of the thick blood films occurring systematically in great malaria control organizations in which the laboratory receives materials from more or less remote localities, particularly in the Brazilian northeast, in regions invaded by Anopheles gambiae. III) An analysis of the causes of failure of the methods of Chorine and Knowles recorded in the literature for such purposes is given, as well as its adaptability for the simultaneous preparation of large numbers of samples. IV) The method is based the protective action of a previous fixation by a dilute solution of formalin, which, without preventing further dehemoglobinization, prevents morphological alterations in the parasites by the action of Knowles solution which is retained in this metod without modification. V) For washing out the acids of the dehemoglobinizating solution as well as for diluting the Giemsa stain, the A. proposes a very simple technique, extremely convenient for such purpose, which consists in adding acetic acid to the distilled water in the proportion of 1 drop for each 10cc of water, and then increasing the hydrogeni-on concentration to pH 7.2 with a 2% sol. of sodium carbonate. As indicator a 0.02% solution purple-bromcresol prepared in accordante to Medalia, is used. In this reaction there is the formation of the acetic acid ↔ sodium acetate, buffer system very suitable for giving a convenient pH and for preventing the precipitation of the dye, which can be used for two or three batches of 700 or 800 slides each, without changing the staining solution. VI) – The method can be summarized as follow: For a small number of samples, Coplin’s or any other staining jar can be used. Large number of slides must be placed in groups of 10 or 15 units each, the slides being separated by a piece of cardboard, according to Barber & Komp. A) Fix in dilute formalin (2%), during 5 minutes. b) Without washing, put in Knowles solution (see the formula in the text), no more than 20 minutes. c)Two successive washings in distilled water, buffered as explained above (which can be used several times). d) Dry and stain with Giemsa solution, prepared by using 1 drop of the stain for each c. c. of buffered distilled water. Time: 1 hour. E) Was in distilled water and dry.