Assessment of BMP responses in an in vitro model of acute ethanol toxicity.

The assay presented here was designed to assess the immediate effects of ethanol (EtOH) exposure on intracellular signaling activated by BMPs (Bone Morphogenetic Proteins). Previous reports of the relationship between EtOH exposure and BMP-dependent signaling have primarily assessed the expression of individual BMPs, changes in BMP target genes or effects on the phosphorylation level of key downstream mediators after days or weeks of in vivo EtOH exposure. What happens to BMP-stimulated signaling immediately following exposure to EtOH remains largely unexplored. Here, the early events of BMP-evoked intracellular signaling were examined in an in vitro model of acute EtOH toxicity. The BMP/Ethanol Stimulation Assay involved first stimulating cultured cells with recombinant BMPs. BMP-evoked intracellular signaling was then allowed to develop for 30 minutes. Next, the cells were exposed to a range of EtOH concentrations for an additional 30 minutes. Finally, the cultures were processed for Western blot analysis or immunofluorescent labeling. This short-term assay: • Permits investigation of EtOH exposure during the initial signaling events downstream of BMP receptor activation • Enables assessment of how the presence of BMPs might protect against cellular injury caused by toxic EtOH levels.

Improving biocatalysis of cefaclor with penicillin acylase immobilized on magnetic nanocrystalline cellulose in deep eutectic solvent based co-solvent.

Deep eutectic solvent (DES), has been considered as a new type of green solvent applied in enzymatic systems. Here, we reported DES-buffer co-solvent as a novel reaction medium for high efficient synthesis of cefaclor by penicilin acylase immobilized on magnetic nanocrystalline cellulose. Effect of DES composition, DES-buffer ratio, temperature, pH, substrate ratio and substrate concentration was systematically investigated. In co-solvent consisting of choline chloride (ChCl):glycol-buffer (7:3, v/v), conversion of 7-ACCA was 94%, synthesis to hydrolysis ratio was 1.8, and yield of cefaclor reached 91%, higher than that in aqueous buffer with optimized yield of 84%, showing the great potential of DES as organic solvent alternative. To the best of our knowledge, this is the first example of biosynthesis of cefaclor in the DES-buffer co-solvent.

Impact of PCV7 vaccination on nasopharyngeal carriage and antimicrobial resistance among children in Turkey.

INTRODUCTION: We aimed to evaluate the effects of 7-valent pneumococcal conjugate vaccine on nasopharyngeal carriage of Streptococcus pneumoniae and antibiotic resistance in children in a well-child clinic in a tertiary children's hospital in Turkey. METHODOLOGY: We collected nasopharyngeal (NP) specimens from 557 two-month-old babies before vaccination. After the study population had received PCV7, NP samples were obtained from 135 babies. Antimicrobial susceptibility testing and serotyping were performed. RESULTS: S. pneumoniae colonized in 48 (8.6%) of the 557 two-month-old babies before vaccination. The follow-up cohort consisted of 135 subjects. The prevalence of PCV7 strain decreased from 33.3% to 19.3% after vaccination. However, non-PCV7 types increased from 66.6% to 80.6% (p = 0.02). Of PCV7 serotypes, 19F was the most frequent serotype before and after vaccination. There was an increase in 6A and 15 of non-PCV7 serotypes after vaccination. Penicillin non-susceptible increased from 56.3% to 80.6% after vaccination (p =0.03). Serotypes 14, 18C, 9V and 6B, which were identified before vaccination, never colonized afterwards. Number of siblings and having sibling with older age of five were determined to be significant effective factors for SP colonization presence after vaccination and antibiotic use was negatively associated with pneumococcal carriage but associated with penicillin non-susceptibility. CONCLUSIONS: Nasopharyngeal carriage rate of S. pneumoniae dropped after PCV7 vaccination, and replacement by NVT pneumococci were also observed. Risk factors for nasopharyngeal carriage included household crowding and having a sibling age five years or older. Penicillin non-susceptibility increased in both VT and NVT strains.

Efeito da penicilina G a cada três semanas sobre o surgimento de Streptococcus viridans resistentes à penicilina na microflora oral / Effect of penicillin G every three weeks on oral microflora by penicillin resistant Viridans Streptococci

FUNDAMENTO: Penicilina G benzatina a cada 3 semanas é o protocolo padrão para a profilaxia secundária para febre reumática recorrente. OBJETIVO: Avaliar o efeito da penicilina G benzatina em Streptococcus sanguinis e Streptococcus oralis em pacientes com doença valvular cardíaca, devido à febre reumática com recebimento de profilaxia secundária. MÉTODOS: Estreptococos orais foram avaliados antes (momento basal) e após 7 dias (7º dia) iniciando-se com penicilina G benzatina em 100 pacientes que receberam profilaxia secundária da febre reumática. Amostras de saliva foram avaliadas para verificar a contagem de colônias e presença de S. sanguinis e S. oralis. Amostras de saliva estimulada pela mastigação foram serialmente diluídas e semeadas em placas sobre agar-sangue de ovelhas seletivo e não seletivo a 5% contendo penicilina G. A identificação da espécie foi realizada com testes bioquímicos convencionais. Concentrações inibitórias mínimas foram determinadas com o Etest. RESULTADOS: Não foram encontradas diferenças estatísticas da presença de S. sanguinis comparando-se o momento basal e o 7º dia (p = 0,62). No entanto, o número existente de culturas positivas de S. oralis no 7º dia após a Penicilina G benzatina apresentou um aumento significativo em relação ao valor basal (p = 0,04). Não houve diferença estatística existente entre o momento basal e o 7º dia sobre o número de S. sanguinis ou S. oralis UFC/mL e concentrações inibitórias medianas. CONCLUSÃO: O presente estudo mostrou que a Penicilina G benzatina a cada 3 semanas não alterou a colonização por S. sanguinis, mas aumentou a colonização de S. oralis no 7º dia de administração. Portanto, a susceptibilidade do Streptococcus sanguinis e Streptococcus oralis à penicilina G não foi modificada durante a rotina de profilaxia secundária da febre reumática utilizando a penicilina G. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0).

BACKGROUND: Benzathine penicillin G every 3 weeks is the standard protocol for secondary prophylaxis for recurrent rheumatic fever. OBJECTIVE: Assess the effect of Benzathine penicillin G on Streptococcus sanguinis and Streptococcus oralis in patients with cardiac valvular disease due to rheumatic fever receiving secondary prophylaxis. METHODS: Oral streptococci were evaluated before (baseline) and after 7 days (day 7) with Benzathine penicillin G in 100 patients receiving routine secondary rheumatic fever prophylaxis. Saliva samples were evaluated for colony count and presence of S. sanguinis and S. oralis. Chewing-stimulated saliva samples were serially diluted and plated onto both nonselective and selective 5% sheep blood agar containing penicillin G. The species were identified using conventional biochemical tests. Minimal inhibitory concentrations were determined with the Etest. RESULTS: No statistical differences were found in the presence of S. sanguinis comparing baseline and day 7 (p = 0.62). However, the existing number of positive cultures of S. oralis on day 7 after Benzathine penicillin G presented a significant increase compared to baseline (p = 0.04). No statistical difference was found between baseline and day 7 concerning the number of S. sanguinis or S. oralis CFU/mL and median minimal inhibitory concentrations. CONCLUSION: This study showed that Benzathine penicillin G every 3 weeks did not change the colonization by S. sanguinis, but increased colonization of S. oralis on day 7 of administration. Therefore, susceptibility of Streptococcus sanguinis and Streptococcus oralis to penicillin G was not modified during the penicillin G routine secondary rheumatic fever prophylaxis. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0).

Implementation of a penicillin allergy skin test / Implementação de um teste cutâneo de alergia à penicilina

The penicillin allergy skin testing is the only accurate and reliable test for penicillin hypersensitivity mediated by IgE. It is useful for identifying patients with doubtful history of allergy. Positive test for major and minor determinants presents a positive predictive value of 50 percent and negative predictive value of 99 percent. In Brazil, the Ministry of Health suggests a protocol for in house made reagents, since they are not commercially available. As the referred protocol does not mention some important details about the test procedures, we propose in the present work to implement them, critically evaluating each step in order to allow the protocol establishment at any health service, with quality and safety.

O teste cutâneo para alergia imediata a penicilina é o único teste validado internacionalmente, sendo que sua grande utilidade reside na avaliação de pacientes com história positiva de alergia a penicilina. O teste positivo para determinantes principais e secundários da penicilina apresenta um valor preditivo positivo de 50 por cento e valor preditivo negativo de 99 por cento. Em nosso meio, o Ministério de Saúde disponibiliza um protocolo para o preparo dos reagentes, uma vez que os mesmos não estão disponíveis comercialmente. Como o referido protocolo não apresenta maiores detalhes sobre o cuidado relativo às etapas de preparo das soluções, bem como faltam algumas considerações no que tange a realização do teste, propusemo-nos no presente trabalho operacionalizar o teste, avaliando de forma crítica e minuciosa cada etapa, de forma que outros profissionais possam reproduzi-lo de maneira mais segura e eficaz.