RESUMO
New promising and versatile materials for the development of in situ sustained release systems consisting of thin films of either poly(2-hydroxyethyl methacrylate) or a copolymer based on poly(ethylene-glycol diacrylate) and acrylic acid were investigated. These polymers were electrosynthesized directly on titanium substrates and loaded with ciprofloxacin (CIP) either during or after the synthesis step. X-ray photoelectron spectroscopy was used to check the CIP entrapment efficiency as well as its surface availability in the hydrogel films, while high-performance liquid chromatography was employed to assess the release property of the films and to quantify the amount of CIP released by the coatings. These systems were then tested to evaluate the in vitro inhibition of methicillin-resistant Staphylococcus aureus (MRSA) growth. Moreover, a model equation is proposed which can easily correlate the diameter of the inhibition haloes with the amount of antibiotic released. Finally, MG63 human osteoblast-like cells were employed to assess the biocompatibility of CIP-modified hydrogel coatings.
Assuntos
Ciprofloxacina/farmacologia , Materiais Revestidos Biocompatíveis/síntese química , Técnicas Eletroquímicas/métodos , Hidrogel de Polietilenoglicol-Dimetacrilato/síntese química , Infecções Relacionadas à Prótese/prevenção & controle , Titânio/efeitos adversos , Antibacterianos/farmacologia , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Humanos , Cinética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microscopia de Fluorescência , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/ultraestrutura , Espectroscopia Fotoeletrônica , Infecções Relacionadas à Prótese/microbiologiaRESUMO
This review describes the ecological, clinical and epidemiological features of emerging vibrios and discusses what laboratory methods are being used for the detection of pathogenic vibrios in clinical, environmental and food samples. After selecting articles illustrative of the current scientific research on pathogenic vibrios, the review focuses on the need for better insight into the risk factors of emerging infections to establish adequate prevention procedures.
Assuntos
Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio/isolamento & purificação , Vibrio/fisiologia , Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Técnicas Bacteriológicas , Biomarcadores/análise , Cólera/etiologia , Cólera/fisiopatologia , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/etiologia , Microbiologia Ambiental , Microbiologia de Alimentos , Gastroenterite/epidemiologia , Gastroenterite/microbiologia , Humanos , Vibrio/patogenicidade , Vibrioses/diagnóstico , Microbiologia da Água , Infecção dos Ferimentos/epidemiologia , Infecção dos Ferimentos/microbiologiaRESUMO
The hepatitis A virus (HAV) is the most common cause of viral infection linked to shellfish consumption. The lack of correlation between the fecal coliform indicators and the presence of enteric viruses in shellfish and their harvesting waters points to the need for molecular methods to detect viruses. We compared two RT-PCR based techniques currently available for the detection of the hepatitis A virus (HAV) in shellfish. Both approaches involve extraction of viral particles by glycine buffer and concentration of virus particles by one or two PEG precipitation steps. One procedure involves as RNA extraction method the use of oligo (dT) cellulose to select poly (A) RNA, and the other uses a system in which total RNA is bound on silica membrane. Comparison of the two RT-PCR based methods highlighted the efficiency of the first approach which is less time-consuming and technically demanding than the second.
Assuntos
Bivalves/virologia , Vírus da Hepatite A/genética , Hepatite A/prevenção & controle , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Frutos do Mar/virologia , Animais , Hepatite A/transmissão , Vírus da Hepatite A/isolamento & purificação , Membranas Artificiais , RNA Mensageiro/análise , RNA Mensageiro/isolamento & purificação , RNA Viral/análise , RNA Viral/isolamento & purificação , Sensibilidade e Especificidade , Dióxido de SilícioRESUMO
A PCR assay for the detection of hepatitis A virus (HAV) in shellfish is described. The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection was accomplished in a single step with the use of primers specific for the VP3-VP1 region of the genome. The procedure detected one 50% tissue culture infective dose (0.6 PFU) per 25 g of shellfish homogenate. Heminested PCR was then carried out to verify the specificity of the PCR products. The method was used to detect HAV in shellfish samples from EU categories B and C and to evaluate the quality of shellfish in routine monitoring for HAV in view of the relevant public health implications of this foodborne disease.