JC polyomavirus nephropathy confirmed by using an in-house polymerase chain reaction method.

We report the case of an isolated JC virus (JCV) infection, without co-infection by polyoma BK virus (BKV), associated with nephropathy 4 years after kidney transplantation. Clinical suspicion followed the observation of a decrease in estimated glomerular filtration rate (eGFR) and a renal allograft biopsy revealing polyomavirus-associated tubulointerstitial nephritis and positivity for SV40. An in-house real-time polymerase chain reaction assay, targeting the presence of JCV and the absence of BKV in biopsy tissue, confirmed diagnosis. Thirteen months after diagnosis, and following therapeutic measures, eGFR remains stable.

The growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities in two rearing systems.

This study evaluated the growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities using two rearing systems. The experiment was conducted in a factorial design (4x2) with four stocking densities (0.1, 0.4, 0.7, and 1.0 g/L), two type of aquarium tanks (glass and ceramic aquariums), and four replicates. The experiment lasted 60 days using 148 juvenile fish (3.05 ± 0.09 g) randomly placed in 32 aquariums (50 L) equipped with filters and aeration. All fish received two meals a day ad libitum (8:00 and 16:00). Water quality parameters such as temperature, dissolved oxygen, pH, and total ammonia were measured. At the end of the experiment, all fish were measured and weighed to determine growth performance and then subjected to parasitological analysis. The data were analyzed with a two-way ANOVA with post-hoc Tukey test (p<0.05). No effects on growth performance at different stocking densities were observed. However, there was an increase in Capillaria pterophylli infestation in the high stocking density within ceramic aquariums. Thus, this study recommends the use of 1.0 g/L for the intensive aquaculture system of freshwater angelfish, and applying cleaning management to avoid parasite infestation, particularly in ceramic aquariums.

The Global Bioequivalence Harmonisation Initiative (GBHI): Report of the fifth international EUFEPS/AAPS conference.

The series of conferences of the Global Bioequivalence Harmonisation Initiative (GBHI) was started in 2015 by the European Federation for Pharmaceutical Sciences (EUFEPS). All GBHI meetings so far were co-organised together with the American Association of Pharmaceutical Scientists (AAPS). Beginning with the 3rd workshop US-FDA joined as co-sponsor - to support global harmonisation of regulatory recommendations for bioequivalence (BE) assessment. At the 5th GBHI conference, the following BE topics were intensively discussed, and the following main conclusions were drawn: (1) Statistical considerations for BE assessment in specific situations covering scaling approaches for highly variable drug (HVD) products, two-stage adaptive design and opportunities of modelling and simulation to support BE: even though special BE study concepts like adaptive designs are not often used in practise so far, a majority of the workshop participants were in favour of a more frequent application of such approaches. The regulatory conditions relevant in this context need further concretisation and harmonisation between the regions. Moreover, modelling and simulation were considered as a promising and evolving approach, also for BE development programmes. (2) Fed versus fasting conditions in BE trials: Findings that BE between generic products could be confirmed only after fasted administration but failed under fed conditions seem more an exception than the rule. Obviously, BCS class IV compounds are most problematic in this context. Differences in critical excipients such as surfactants or pH-modifiers may be relevant reasons for different sensitivity for interactions in fasted versus fed conditions. Consequently, such deviations in composition of generic preparations should be avoided. Moreover, confirmation of BE may be generally difficult comparing different dosage forms, such like capsules versus tablets, especially in fed state. (3) BE assessment of locally acting drug products applied topically to the skin: Appropriateness and potential benefit of in-vitro tests as alternatives to clinical efficacy studies have been comprehensively discussed. In addition to the already well-established in-vitro release and permeation tests, other techniques were suggested, e.g., Raman spectroscopy or dermal open flow microperfusion. Validation of those methods is challenging and, despite significant progress already achieved during previous years, more research is needed before they may be fully accepted for regulatory purposes. (4) BE evaluation of narrow therapeutic index (NTI) drugs: The discrepancies amongst regulatory agencies in necessity of tighter BE acceptance ranges, the recommendations for inclusion of peak and total drug exposure into BE assessment with more restrictive criteria and the importance of comparison of the product-related within-subject variability for NTI drugs were debated. Arguments in favour and against the different approaches were presented and discussed but need further consideration before harmonisation can be achieved. The highly interactive meeting and extensive exchange between regulators and scientists from industry and academia resulted in useful progress in open BE issues and supported the goal of science-driven harmonisation.

Prevalence of human cytomegalovirus congenital infection in Portuguese newborns.

Human cytomegalovirus (HCMV) is considered the most frequent cause of congenital infection, occurring in 0.2 to 2.2% of all live births. Since this is a wide range of prevalences observed in different studies, it would be desirable to investigate the prevalence of this infection at national level. The aim of this study was the evaluation of the national prevalence of HCMV congenital infection. We analysed a total of 3,600 Guthrie cards collected from Portuguese newborns during a period of 14 months (August 2003 to September 2004). The cards covered all regions of Portugal and were proportional to the number of births in each region. A heat DNA extraction method was used, followed by DNA amplification by nested PCR. Sensitivity and specificity of this method were evaluated as 93% and 100%, respectively, using 28 cards from HCMV-positive and 280 cards from HCMV-negative children. The national prevalence of congenital HCMV was determined as 1.05% (95% confidence interval: 0.748-1.446). This is the first study of the prevalence of HCMV congenital infection at national level in Portugal. It suggests that Portugal may have one of the highest prevalences of congenital HCMV infection in Europe.

Implementing the additional strength biowaiver for generics: EMA recommended approaches and challenges for a US-FDA submission.

This review describes the EMA requirements on biowaivers for additional strengths of immediate release and modified release oral solid dosage forms focused on generic applications and highlights the challenges for a simultaneous EMA and FDA submission. Some specificities of the current EMA guidelines are compared with the current FDA Guidance for Industry, with a special focus on the strength to be investigated in vivo, formulation suitability for biowaiver, and optimizing dissolution studies for additional strength biowaivers. In Europe, the same principles applied for generics may be considered for deriving the biowaivers for innovator products. Several case studies are presented to illustrate the challenges of applying for additional strength biowaivers in EMA and FDA simultaneously.

Lawsonella clevelandensis as the causative agent of a breast abscess.

Lawsonella clevelandensis is a Gram-stain-positive, partially acid-fast, anaerobic, being considered a new species within a new genus in the suborder Corynebacterineae. There are only a few cases reported worldwide. This is a fastidious microorganism, difficult to identify by conventional methods, leading to inappropriate treatments. The authors report a case of a 29-year-old woman with a 3-week evolution of a breast nodule. There was a family history of breast carcinoma. Samples were collected for histological and microbiological examination. Gram staining revealed Gram-positive filamentous bacilli, acid-fast-positive. The cultural examination revealed a Lawsonella clevelandensis that was confirmed by molecular methods. At the last follow up, the evolution was favorable; the abscess was resolved, with no evidence of recurrence. To our knowledge the present case was the first reported in Europe.

Implementing the Biopharmaceutics Classification System in Drug Development: Reconciling Similarities, Differences, and Shared Challenges in the EMA and US-FDA-Recommended Approaches.

The US-FDA recently posted a draft guideline for industry recommending procedures necessary to obtain a biowaiver for immediate-release oral dosage forms based on the Biopharmaceutics Classification System (BCS). This review compares the present FDA BCS biowaiver approach, with the existing European Medicines Agency (EMA) approach, with an emphasis on similarities, difficulties, and shared challenges. Some specifics of the current EMA BCS guideline are compared with those in the recently published draft US-FDA BCS guideline. In particular, similarities and differences in the EMA versus US-FDA approaches to establishing drug solubility, permeability, dissolution, and formulation suitability for BCS biowaiver are critically reviewed. Several case studies are presented to illustrate the (i) challenges of applying for BCS biowaivers for global registration in the face of differences in the EMA and US-FDA BCS biowaiver criteria, as well as (ii) challenges inherent in applying for BCS class I or III designation and common to both jurisdictions.

Growth and survival of the native oyster Crassostrea gasar cultured under different stocking densities in two grow-out systems in tropical climate / [Crescimento e sobrevivência da ostra nativa Crassostrea gasar cultivada em diferentes densidades em dois sistemas em clima tropical]

Survival and growth of the native oyster Crassostrea gasar along the juvenile and adult phases were evaluated in three different stocking densities [low (D), medium (2D) and high (3D)] and in two grow-out systems (fixed and floating system). The fixed system consisted of a rack made with PVC, fixed from the bottom with wood sticks. The floating system consisted of floating bags suspended by a rack made with PVC and maintained submerged from the seawater surface by eight floats. Survival and shell height of oysters cultured after 30, 60 and 90 days were registered in each phase and in each grow-out system. Results showed that the grow-out system did not affect survival and growth of C. gasar in the juvenile and adult phases. The tested densities affected the survival of oysters cultured over time in both phases but did not affect oyster growth. At times analyzed, it was observed positive growth in juvenile oysters grow after 90 days of culture. However, in the adult phase, no growth was observed after 90 days of culture. Oyster yield was higher in the density 3D, in both juvenile and adult phases. These findings contributed to the development of the oyster C. gasar culture.(AU)

A sobrevivência e o crescimento da ostra nativa Crassostrea gasar nas fases juvenil e adulta foram avaliados sob três diferentes densidades de estocagem [baixa (D), média (2D) e alta (3D)] e dois sistemas de engorda (fixo e flutuante). O sistema fixo consistiu em uma mesa de PVC, fixada na parte inferior com varas de madeira. O sistema flutuante consistiu em travesseiros flutuantes suspensos por uma mesa de PVC e mantidas submersas da superfície da água do mar por oito flutuadores. Registraram-se sobrevivência e altura da concha de ostras cultivadas após 30, 60 e 90 dias, em cada fase (juvenil e adulta) e em cada sistema (fixo e flutuante). Os resultados mostraram que o sistema de engorda não afetou a sobrevivência e o crescimento de C. gasar nas fases juvenil e adulta. As densidades testadas afetaram a sobrevivência das ostras ao longo do tempo, em ambas as fases, mas não afetaram o crescimento em altura. Nos tempos analisados, ostras juvenis apresentaram crescimento após 90 dias de cultivo. Porém, na fase adulta, não foi observado crescimento após 90 dias de cultivo. A produção de ostras, foi maior na densidade 3D, nas fases juvenil e adulta. Os presentes achados contribuíram para o desenvolvimento do cultivo da ostra C. gasar.(AU)

Sanitary conditions of the Goliath grouper Epinephelus itajara in captivity on estuarine environment: case report / Condições sanitárias do mero Epinephelus itajara em cativeiro em ambiente estuarino: relato de caso

This study aimed to report the sanitary conditions through the hematological analysis of grouper E. itajara reared in captivity on estuarine conditions. Seven Goliath groupers (1,881.5±1,246.03g) were captured and kept in two tanks located on estuary. After 20 days, fish were collected for morphologic and hemato-physiologic evaluation. Two fish had clinical signs such as hemorrhagic spots and loss of scale due to agonistic behavior. Blood samples were collected, and the hematological parameters (biochemical, erythrogram and leukogram) were determined. Blood cells were characterized by their size, color and shape. Univariate statistic and principal components analysis were used to identify a hematological standard between fish with or without clinical signs. Four leukocyte types were found: lymphocyte, monocyte, neutrophil and basophil. Regardless of the clinical signs the cell morphology did not present any difference among the fish. However, there is a significant correlation between erythrocyte and lactate on fish with clinical signs. Thus, agonistics encountered among the fish is a stressing factor in captivity conditions making it necessary to have adequate management related to the size of fish and stocking density.(AU)

Este estudo avaliou as condições sanitárias do peixe mero E. itajara mantido sob cativeiro, em condições estuarinas, pelas análises hematológica e morfológica. Sete peixes (1.881,5±1.246,03g) foram capturados e mantidos em tanques localizados no estuário. Após 20 dias, os peixes foram coletados para avaliações morfológica e hematológica. Dois peixes tiveram sinais clínicos, como manchas hemorrágicas e perda de escamas devido ao comportamento agonístico entre os peixes. Amostras sanguíneas foram coletadas dos peixes anestesiados com auxílios de seringas umedecidas com EDTA 3%. Determinaram-se os parâmetros hematológicos (bioquímico, eritrograma e leucograma). Células sanguíneas foram caracterizadas por seu tamanho, cor e padrão. Estatística univariada e análises de componentes principais foram usadas para identificar um padrão hematológico entre os peixes com e sem sinais clínicos. Quatro tipos de leucócitos foram encontrados: linfócito, monócito, neutrófilo e basófilo. Os sinais clínicos não apresentaram diferença entre os peixes e a morfologia celular. Contudo, observou-se uma correlação entre os peixes com sinais clínicos e a quantidade de eritrócitos e lactato. Assim, encontros antagônicos entre os peixes são um fator estressante em condições de cativeiro, tornando-se necessário um manejo mais adequado relacionado ao tamanho dos indivíduos e à densidade de estocagem.(AU)

Simple method for determination of paraquat in plasma and serum of human patients by high-performance liquid chromatography.

A simple and fast HPLC system is presented for quantifying paraquat in human plasma and serum using 1,1'-diethyl-4,4'-bipyridyldiylium (diethyl paraquat) as an internal standard. An octadecyl-silica column is used with an eluent of 10% acetonitrile (v/v) containing sodium 1-octanesulphonic acid (3.0 mM) and a diethylamine-orthophosphoric acid buffer (pH 3). Unlike with other techniques, sample treatment requires only the precipitation of protein contents by 6% perchloric acid (v/v) in methanol. The method has a limit of detection of 0.1 microg/ml and is linear up to 10 microg/ml. The serum of four patients and the plasma of one patient with paraquat intoxication's were analysed and positive identification and quantification was readily achieved. One of those patients survived, partially given the rapid disclosure of his levels of paraquat. Therefore, this method is suitable for quantification of paraquat in toxicological samples. It may be used as a prognostic tool in critical case detoxification and to quickly identify potentially salvageable patients for enrollment in new hemofiltration studies.

Metabolism of primaquine by liver homogenate fractions. Evidence for monoamine oxidase and cytochrome P450 involvement in the oxidative deamination of primaquine to carboxyprimaquine.

The role of monoamine oxidase (MAO) and cytochrome P450 (P450) in the oxidative deamination of primaquine by rat liver fractions was studied. Rat liver fractions including liver homogenate, mitochondria, microsomes and 100,000 g supematant fractions were prepared from a pool of rat livers and characterised using benzylamine as a probe for MAO activity and N,N-dimethylbenzamide as a probe for P450 N-dealkylation activity. Incubation of all fractions with primaquine yielded carboxyprimaquine as the only metabolite detectable by HPLC. The mitochondrial fraction, which contained MAO activity but not P450 activity, presented the highest Vmax/K(M) value for the formation of carboxyprimaquine (8.5 x 10(-6) dm3mg(-1)h(-1). A substantially lower Vmax/K(M) value (1.3 x 10(-6) dm3mg(-1)h(-1)) was obtained in the microsomal fraction, which contained P450 but not MAO activity. The liver homogenate fraction presented a similar value (1.8 x 10(-6) dm3mg(-1)h(-1), though it contained both enzyme systems. Incubations of all the fractions that presented MAO activity, in presence of the MAO inhibitor pargiline, resulted in a marked inhibition of primaquine oxidation. P450 inhibitor SKF 525-A effectively inhibited primaquine metabolism in the microsomal fraction but inhibition in the liver homogenate was less effective. The results are consistent with an important role for MAO in primaquine biotransformation, though clearly metabolism by P450 has a contribution role.

[Infectious endocarditis caused by Q fever. Apropos of a clinical case]. / Endocardite infecciosa a febre Q. A propósito de um caso clínico.

Endocarditis is a rare, but some times fatal, complication of Q fever. Its diagnosis is difficult and it is based on non-specific cardiac findings and a high title of phase I antibodies. The treatment is based on tetracyclines alone or in combination with cotrimoxazole, for long periods of time. The therapeutic efficacy is evaluated by the measurement of phase I antibodies, every three months. The relapses are frequent despite the long period of antibiotic therapy. We report what is probably the first case of Q fever prosthesis endocarditis in Portugal, as a complication following an acute episode of Q fever.

Environment and Health in Children Day Care Centres (ENVIRH) - Study rationale and protocol.

BACKGROUND: Indoor air quality (IAQ) is considered an important determinant of human health. The association between exposure to volatile organic compounds, particulate matter, house dust mite, molds and bacteria in day care centers (DCC) is not completely clear. The aim of this project was to study these effects. METHODS ­ STUDY DESIGN: This study comprised two phases. Phase I included an evaluation of 45 DCCs (25 from Lisbon and 20 from Oporto, targeting 5161 children). In this phase, building characteristics, indoor CO2 and air temperature/relative humidity, were assessed. A children's respiratory health questionnaire derived from the ISAAC (International Study on Asthma and Allergies in Children) was also distributed. Phase II encompassed two evaluations and included 20 DCCs selected from phase I after a cluster analysis (11 from Lisbon and 9 from Oporto, targeting 2287 children). In this phase, data on ventilation, IAQ, thermal comfort parameters, respiratory and allergic health, airway inflammation biomarkers, respiratory virus infection patterns and parental and child stress were collected. RESULTS: In Phase I, building characteristics, occupant behavior and ventilation surrogates were collected from all DCCs. The response rate of the questionnaire was 61.7% (3186 children).Phase II included 1221 children. Association results between DCC characteristics, IAQ and health outcomes will be provided in order to support recommendations on IAQ and children's health. A building ventilation model will also be developed. DISCUSSION: This paper outlines methods that might be implemented by other investigators conducting studies on the association between respiratory health and indoor air quality at DCC.

ANTECEDENTES: A qualidade do ar interior (IAQ) é considerada um determinante importante da saúde humana. A associação entre a exposição a compostos orgânicos voláteis, partículas, ácaros, bolores e bactérias em creches (DCC) não é perfeitamente clara. O objectivo deste estudo foi estudar esses efeitos. METODOLOGIA ­ DESENHO DO ESTUDO: Este estudo decorreu em duas Fases. A Fase I incluiu uma avaliação de 45 DCCs (25 em Lisboa e 20 no Porto, visando 5.161 crianças). Nesta Fase, foram avaliadas as características dos edifícios, o CO2 e a temperatura ambiente/humidade relativa no interior. Também foi distribuído um questionário de saúde respiratória das crianças derivado do ISAAC (Estudo Internacional sobre a Asma e Alergias em Crianças). A Fase II englobou duas avaliações e incluiu 20 DCCs seleccionadas da fase I, após uma análise de clusters (11 em Lisboa e 9 no Porto, visando 2.287 crianças). Nesta Fase, foram recolhidos dados sobre a ventilação, IAQ (qualidade do ar interior), parâmetros de conforto térmico, saúde em termos respiratórios e alérgicos, marcadores biológicos de inflamação das vias respiratórias, padrões de infecção de vírus respiratórios e stress dos pais e crianças. RESULTADOS: Na Fase I, foram recolhidas as características dos edifícios, o comportamento dos ocupantes e os marcadores de ventilação, para todas as DCCs. A taxa de resposta ao questionário foi de 61,7% (3.186 crianças).A Fase II incluiu 1.221 crianças. Os resultados da associação entre as características dos DCC, a IAQ e os resultados para a saúde serão fornecidos para apoiar as recomendações sobre a IAQ e a saúde das crianças. Um modelo de ventilação de edifícios também será desenvolvido. DISCUSSÃO: Este documento destaca a metodologia que pode ser implementada por outros investigadores que realizam estudos sobre a associação entre a saúde respiratória e a qualidade do ar interior, em creches e infantários.

Environment and Health in Children Day Care Centres (ENVIRH) - Study rationale and protocol.

BACKGROUND: Indoor air quality (IAQ) is considered an important determinant of human health. The association between exposure to volatile organic compounds, particulate matter, house dust mite, molds and bacteria in day care centers (DCC) is not completely clear. The aim of this project was to study these effects. METHODS - STUDY DESIGN: This study comprised two phases. Phase I included an evaluation of 45 DCCs (25 from Lisbon and 20 from Oporto, targeting 5161 children). In this phase, building characteristics, indoor CO2 and air temperature/relative humidity, were assessed. A children's respiratory health questionnaire derived from the ISAAC (International Study on Asthma and Allergies in Children) was also distributed. Phase II encompassed two evaluations and included 20 DCCs selected from phase I after a cluster analysis (11 from Lisbon and 9 from Oporto, targeting 2287 children). In this phase, data on ventilation, IAQ, thermal comfort parameters, respiratory and allergic health, airway inflammation biomarkers, respiratory virus infection patterns and parental and child stress were collected. RESULTS: In Phase I, building characteristics, occupant behavior and ventilation surrogates were collected from all DCCs. The response rate of the questionnaire was 61.7% (3186 children). Phase II included 1221 children. Association results between DCC characteristics, IAQ and health outcomes will be provided in order to support recommendations on IAQ and children's health. A building ventilation model will also be developed. DISCUSSION: This paper outlines methods that might be implemented by other investigators conducting studies on the association between respiratory health and indoor air quality at DCC.

Fauna parasitária e relação parasito-hospedeiro de tambaquis criados na região do Baixo São Francisco, nordeste do Brasil / Parasitic fauna and parasite-host relationship of tambaqui reared in São Francisco river basin, Brazilian northeast

Este trabalho teve como objetivo avaliar a fauna parasitária de tambaquis na região do Baixo São Francisco-AL/SE-Brasil e correlacionar os índices de prevalência e intensidade média com fatores bióticos e abióticos. Foram coletados 252 espécimes para análise parasitológica de 10 pisciculturas. Os parasitos foram contabilizados, identificados, e determinaram-se os índices de prevalência e intensidade média, que foram correlacionados com fatores bióticos e abióticos. Dos peixes coletados, 65,5% estavam parasitados por pelo menos um táxon. Foram encontrados 10 táxons: Monogeneas, Ichthyophthirius multifiliis, tricodinídeos, Piscinoodinium pillulare, Ichthyobodo sp., Dolops carvalhoi, Lernaea cyprinacea, Procamallanus (Spirocamallanus) inopinatus, Henneguya sp. e Myxobolus sp. As maiores prevalências foram encontradas para Monogeneas (49,2%) e Myxobolus sp. (31,5%). Correlações negativas entre prevalência e fatores bióticos (peso e comprimento) foram observadas para Monogeneas (r2= -0,49; r2= -0,43), Myxobolus sp. (r²= -0,46; r²= -0,39) e Henneguya sp. (r²= -0,41; r²= -0,39). O fator abiótico temperatura apresentou correlação negativa com as prevalências de Lernaea cyprinacea (r= -0,39) e tricodinídeos (r= -0,33), enquanto a condutividade elétrica apresentou correlação positiva (r= 0,40) com a prevalência de tricodinídeos. Conclui-se que a fauna parasitária dos tambaquis cultivados na região do Baixo São Francisco é diversificada e com a carga parasitária dependente da qualidade de água e do estágio de desenvolvimento dos peixes.(AU)

This study investigated the parasitic fauna of tambaquis reared in lower Sao Francisco region-Al/SE-Brazil correlating parasitic indices to abiotic and biotic factors. A total of 252 specimens of tambaqui were collected in ten fish farms for parasitological analysis. The parasites were counted, identified and the parasitological indices were determined and correlated to biotic and abiotic factors. Of all collected fish, 65,5 % were parasitized by at least one taxon. Ten taxa were found: Monogeneans, Ichthyophthirius multifiliis, trichodinids Piscinoodinium pillulare, Ichthyobodo sp, Dolops carvalhoi, Lernaea cyprinacea, Procamallanus (Spirocamallanus) inopinatus, Henneguya sp. and Myxobolus sp. The higher prevalences were found to monogeneans (49.2%) and Myxobolus sp. (31.5%). Negative correlation of prevalence and biotic factor (weight and length) were observed to monogeneans (r 2 = -0.49, r 2 = -0.43), Myxobolus sp (r²= -0.46; r²= -0.39) and Henneguya sp (r²= -0.41; r²= -0.39). Abiotic factor of temperature presented a negative correlation to prevalence of Lernaea cyprinacea and trichodinids (r= -0.39 e r= -0.33, respectively) and the electric conductivity presented positive correlation to trichodinids (r= 0.40). It was concluded that parasitic fauna of tambaquis cultured in Lower São Francisco region is diversified and the parasitic load dependent on water parameters and fish growth.(AU)

Isolation of a fusion protein containing the antigenic domain 1 of human cytomegalovirus glycoprotein B and its application in ELISA tests.

The glycoprotein B (gB) of human cytomegalovirus represents a dominant antigen for the humoral immune response. The immunodominant region on gB is the antigenic domain 1 (AD-1), a complex structure that requires a minimal continuous sequence of more than 75 amino acids for antibody binding. In this study, this domain was expressed in Escherichia coli as a fusion protein with beta-galactosidase but yielded insoluble protein aggregates as inclusion bodies. To recover the fusion protein, inclusion bodies were solubilized by two extractions with urea 8 M: and the fusion protein then isolated using gel filtration chromatography. After confirmation of fusion protein antigenicity by Western blotting, the purified product was used as the capturing antigen in an enzyme-linked immunosorbent assay (ELISA) to determine the presence of viral antibodies in serum samples of pregnant women. A cut-off point of approximately 0.2 absorbance units could discriminate the results of seropositive from seronegative pregnant women. The data indicates the potential usefulness of the fusion protein for the development of immunoassay for detection of the HCMV antibodies.

Aleitamento materno - Nivel socio-economico medio e baixo. / Breast feeding - Median and low socioeconomic level

Na estatistica apresentada neste trabalho observamos um baixo indice de aleitamento materno nas classes de medio e baixo nivel socio-economico na regiao pesquisada, bem como os motivos desde baixo indice de algumas sugestoes no sentido de eleva-lo. Aproveitamos ainda para ratificar uma vez mais as vantagens do leite materno, para o bom desenvolvimento da crianca