This work contributes in vaccines down-stream process by introducing a novel platform for in-vitro monitoring of vaccine-adjuvant delivery profile as a crucial preclinical optimizing step in mucosal vaccines. Nano and micro particles of Calcium phosphate (Cap) vaccine-adjuvant were encapsulated in Chitosan and Alginate polymeric carriers. Adjuvants release profiles monitored in a permeable bag at 37°C, pH 2, incubated in isotonic buffer for 96 hours. The released Calcium in the outer buffer was monitored and compared in-addition to the carrier’s swelling and biophysical properties. The adjuvants and carriers did not interfere with the proliferation of cultured hepatocytes an indicator of their safe use; Chitosan’s viscosity and swelling were higher than Alginate. Chitosan’s Zeta-potential was significantly high positive, while Cap and Alginate were negative. The prepared CaP and Chitosan particles were in nano-size, while the ready-made CaP adjuvant and Alginate were in micro-size using zeta-seizer and scanning electron-micrograph. The release of nano-size particle was in ascending, extended and controlled manner compared to micro-size adjuvant. Moreover, nano-adjuvant release profile from Chitosan was superior compared to Alginate. The core controlling factors in vaccine-adjuvant sustained release includes; smaller adjuvant particles (nano-size), carrier’s low swelling, high viscosity and importantly carrier-adjuvant entrapment reversibility. Chitosan offers sustained ascending superior capacity in releasing Nano-Cap adjuvant. This novel in-vitro pre-clinical study answer a crucial downstream preparative step for optimizing mucosal vaccines before their direct routine in-vivo trial on animal regardless of adjuvant’s particle size or delivery kinetics.

Este trabajo contribuye a la investigación de vacunas, a través de una plataforma in vitro que monitorea los perfiles de liberación vacuna-adyuvante, como paso crucial para el desarrollo preclínico de vacunas mucosales. Las nano y macropartículas de fosfato de calcio (Cap), se encapsularon en sistemas de liberación de quitosana y alginato. El perfil de liberación del adyuvante fue monitoreado en membranas permeables a 37ºC, pH2 e incubado en tampón isotónico por 96 horas. Se monitoreó el calcio liberado en el tampón externo y se comparó con la capacidad de hidratación del sistema de liberación utilizado y sus características biofísicas. Los adyuvantes y sistemas de liberación no interfirieron con la proliferación de cultivos de hepatocitos, demostrando un uso seguro. La viscosidad de la quitosana y su nivel de hidratación fueron mayores que los del alginato, mientras que el potencial zeta de la quitosana fue altamente positivo y el del alginato negativo. Las formulaciones de Cap y las partículas de quitosana tenían tallas nanométricas, mientras que el Cap en alginato formó micropartículas que se observaron en zeta seizer y microscopios electrónicos de barrido. El perfil de liberación de las nanopartículas ocurrió de forma ascendente, extendida y controlada en comparación con el de las micropartículas. Además, el perfil de liberación de la quitosana fue superior al del alginato. Los factores esenciales a controlar en sistemas de liberación con capacidad adyuvante incluyen: partículas adyuvantes de pequeño tamaño (nano), sistemas de liberación con bajo perfil de hidratación, alta viscosidad y poder de encapsulación reversible. La quitosana ofrece una capacidad superior para la liberación del adyuvante nano-Cap. Este novedoso estudio, responde a la necesidad de optimizar las formulaciones antes de los estudios in vivo en animales, sin tener en cuenta el tamaño de partículas o la cinética de distribución.

Although analysis of toxin-antitoxin (TA) systems can be instructive, to date, there is no information on the prevalence and identity of TA systems based on a large panel of Acinetobacter baumannii clinical isolates. The aim of the current study was to screen for functional TA systems among clinical isolates of A. baumannii and to identify the systems’ locations. For this purpose, we screened 85 A. baumannii isolates collected from different clinical sources for the presence of the mazEF, relBE and higBA TA genes. The results revealed that the genes coding for the mazEF TA system were commonly present in all clinical isolates of A. baumannii. Reverse transcriptase-polymerase chain reaction analysis showed that transcripts were produced in the clinical isolates. Our findings showed that TA genes are prevalent, harboured by chromosomes and transcribed within A. baumannii. Hence, activation of the toxin proteins in the mazEF TA system should be investigated further as an effective antibacterial strategy against this bacterium.

CONTEXTO E OBJETIVOS: As bactérias resistentes estão surgindo em todo o mundo como uma ameaça ao resultado favorável no tratamento de infecções comuns em ambientes comunitários e hospitalares. O presente estudo foi realizado com o objetivo de avaliar a incidência de Klebsiella pneumoniae produtoras de beta-lactamases de espectro estendido (BLEEs) em pacientes com infecção do trato urinário em diferentes estações do ano, para verificar a prevalência dos genes blaTEM, blaSHV e blaCTX-M responsáveis para a produção de BLEEs em K. pneumoniae productoras de BLEEs, em três cidades do Irã, e investigar o perfil de sensibilidade antimicrobiana de K. pneumoniae em diferentes estações. DESENHO E LOCAL: Estudo retrospectivo realizado em pacientes com infecções do trato urinário em cinco hospitais no Irã. MÉTODO: Duzentos e oitenta e oito isolados clínicos de K. penumoniae foram coletados entre março de 2007 e abril de 2008 em cinco hospitais de três cidades no Irã. BLEEs foram identificados por métodos fenotípicos e genotípicos. K. pneumoniae produtoras de BLEEs foram avaliadas contra antibióticos não beta-lactâmicos. Os genes codificadores de BLEEs (blaSHV, TEM e CTX-M) foram investigados. RESULTADOS: Entre os 288 isolados clínicos de K. pneumoniae, 37,7%, 46,7% e 15,6% eram provenientes dos hospitais em Ilam, Tehran e Tabriz, respectivamente, dos quais 39,4%, 50,7% e 45,8% eram K. pneumoniae produtoras de BLEEs nos hospitais em Ilam, Milad e Eman Reza, respectivamente. CONCLUSÃO: De acordo com os resultados deste estudo, a resistência às cefalosporinas de terceira geração é maior nos meses frios do que nos meses quentes.

CONTEXT AND OBJECTIVES: Resistant bacteria are emerging worldwide as a threat to favorable outcomes from treating common infections in community and hospital settings. The present investigation was carried out to study the incidence of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae in patients with urinary tract infection in different seasons of the year, in order to determine the prevalence of the genes blaTEM, blaSHV and blaCTX-M, which are responsible for ESBL production among ESBL-producing K. pneumoniae, in three cities in Iran, and to investigate the antimicrobial susceptibility pattern of K. pneumoniae in different seasons. DESIGN AND SETTING: Retrospective study carried out among patients with urinary tract infections in five hospitals in Iran. METHOD: Two hundred and eighty-eight clinical isolates of K. pneumoniae were collected between March 2007 and April 2008 from five hospitals in three cities in Iran. ESBLs were identified by phenotypic and genotypic methods. ESBL-producing Klebsiella pneumoniae were evaluated against non-beta-lactam antibiotics. Genes coding for ESBLs (blaSHV, TEM and CTX-M) were screened. RESULTS: Among the 288 clinical isolates of K. pneumoniae, 37.7%, 46.7% and 15.6% were obtained from hospitals in Ilam, Tehran and Tabriz, respectively, of which 39.4%, 50.7% and 45.8% were ESBL-producing K. pneumoniae in Ilam, Milad and Emam Reza hospitals, respectively. CONCLUSION: According to the results from this study, resistance to third-generation cephalosporins is higher during the cold months than during the warm months.