Subject(s)
Animals , Rats , Bacterial Proteins/biosynthesis , Klebsiella Infections/drug therapy , Klebsiella pneumoniae/enzymology , Sepsis/drug therapy , beta-Lactamases/biosynthesis , Drug Therapy, Combination/methods , Gentamicins/therapeutic use , Klebsiella Infections/microbiology , Minocycline/analogs & derivatives , Minocycline/therapeutic use , Polymyxins/therapeutic use , Thienamycins/therapeutic useABSTRACT
BACKGROUND: Animal models are useful to evaluate the efficacy of antimicrobials in experimental sepsis. AIM: To elucidate the steps of producing an experimental model for the treatment of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae sepsis METHODS: Several ESBL inoculums ranging from 1.5x109 colony-forming units per milliliter (CFU/mL) to 2.0x1010 CFU/mL were administered by peritoneal injection in adults Wistar rats. Outcomes and microbiological data of quantitative peritoneal and blood cultures were observed in untreated animals. Animals which received 2.0x1010 CFU/mL inoculums were treated with single meropenem dose (30mg/kg) after one hour and those which received 1.0x1010 CFU/mL inoculums were treated immediately with three doses of meropenem 50 mg/kg. Outcomes were observed for 24 hours after inoculation. RESULTS: Solutions with 1.5 x109 and 6.0x109 CFU/mL were not lethal within 24 hours. Inoculums of 1.0x1010 CFU/mL were lethal in 80% and solutions with 2.0x1010 CFU/mL were lethal in 100% of animals. ESBL lethal sepsis (1.0x1010CFU/mL) was treated immediately with 50 mg/kg of meropenem every eight hours for 24 hours and presented 40% mortality compared with 80% mortality of the control group (p=0.033). Quantitative cultures of peritoneal fluid presented 104 CFU/mL or less for treated animals compared to more than 105 for untreated animals (p=0.001). CONCLUSION: Inoculums of 1.0x1010CFU/mL achieved the best results to study a model of lethal sepsis and this model of treatment of carbapenem-susceptible Enterobacteriaceae can serve as control to further evaluation of treatment of carbapenemase-producing Enterobacteriaceae models. .
RACIONAL: Modelos animais são importantes para avaliar a eficácia de antimicrobianos e a validação do sítio de infecção e a carga bacteriana. OBJETIVO: Definir a concentração do inóculo bacteriano, a dose e o tempo de administração de antimicrobianos a fim de validar um modelo experimental para o tratamento de Klebsiella pneumoniae produtora de betalactamase de amplo espectro em sepse letal. MÉTODO: Inóculos de Klebsiella pneumoniae produtora de betalactamase de espectro estendido de 1,5x109 unidades formadoras de colônias por mililitro (UFC/ml) a 2,0x1010 UFC/ml foram administrados via injeção peritoneal em ratos Wistar adultos. Sobrevida e dados microbiológicos de hemoculturas e culturas quantitativas de fluido peritoneal foram avaliados inicialmente em animais não tratados. Animais inoculados com 2,0x1010 UFC/ml foram tratados dose única de meropenem (30mg/kg) e animais inoculados com 1,0x1010 UFC/ml foram tratados imediatamente com meropenem (50 mg/kg) por 24 horas e os desfechos foram avaliados após 24 horas da inoculação. RESULTADOS: Soluções com 1,5 x109 e 6,0x109 UFC/ml não foram letais. Inóculos de 1,0x1010 UFC/ml e de 2,0x1010UFC/ml foram letais em 80% e 100% dos animais respectivamente. Sepse letal (1.0x1010CFU/mL) com tratamento imediato e por 24 horas apresentou 40% de mortalidade, comparada com 80% nos controles (p=0.033). Culturas quantitativas de fluido peritoneal apresentaram ≤104 UFC/ml enquanto que controles sem tratamento apresentaram >105 UFC/ml (p=0.001). CONCLUSÃO: Modelo experimental com inóculo de 1,0x1010UFC/ml submetido ao tratamento imediato e por 24 horas foi capaz de avaliar resposta microbiológica e de sobrevida podendo ser modelo de embasamento e de controle para tratamento de sepse letal por Klebsiella pneumoniae produtora de carbapenemase. .
Subject(s)
Animals , Female , Male , Rats , Anti-Infective Agents/therapeutic use , Disease Models, Animal , Klebsiella Infections/drug therapy , Klebsiella pneumoniae/enzymology , Sepsis/drug therapy , beta-Lactamases/biosynthesis , Rats, WistarABSTRACT
Antiretroviral therapy (ART) has reduced morbidity and mortality related to human immunodeficiency virus (HIV) infection, but in spite of this advance, HIV mutations decrease antiretroviral susceptibility, thus contributing to treatment failure in patients. Genotyping HIV-1 allows the selection of new drugs after initial drug failure. This study evaluated the genotypic profile of HIV-1 isolates from treated (drug-experienced) patients in Paraná, Brazil. The prevalence of mutations in reverse transcriptase (RT) and protease (PR) genes were assessed. We analyzed 467 genotypes of patients with HIV-1 viral loads above 1,000 copies/mL. Mutations at HIV-1 RT and PR genes and previously used ART regimens were recorded. The most prevalent RT mutations were: 184V (68.31 percent), 215YF (51.6 percent), 103NS (46 percent), 41L (39.4 percent), 67N (38.54 percent), 210W (23.5 percent), 190ASE (23.2 percent), and 181C (17.4 percent). PR mutations were 90M (33.33 percent), 82ATFS (29 percent), 46I (26.8 percent) and 54V (22.2 percent). The prevalence of mutations was in line with previous national and international reports, except to nonnucleoside analogue reverse transcriptase inhibitors related mutations, which were more prevalent in this study. Previous exposure to antiretroviral drugs was associated with genotypic resistance to specific drugs, leading to treatment failure in HIV patients.
Subject(s)
Adult , Female , Humans , Male , Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , HIV Protease/genetics , HIV Reverse Transcriptase/genetics , HIV-1 , Mutation/genetics , Antiretroviral Therapy, Highly Active , Brazil , Drug Resistance, Viral/genetics , Genotype , HIV Infections/virology , HIV-1 , Treatment Failure , Viral LoadABSTRACT
OBJECTIVE: Because epidemiological data on circulating HIV subtypes among HIV-positive patients in the state of Paraná were not known until now, the aims of this study were to describe the genetic diversity profile of HIV-1 in treated patients in Paraná, Brazil, and report the differences in protease (PR) and reverse transcriptase (RT) mutations in HIV-1 subtypes. PATIENTS AND METHODS: A cross-sectional study was conducted from 2003 to 2006. Plasma viral RNA of 389 patients was extracted and PR and RT genes were polymerase chain reaction-amplified and sequenced. Sequences were subtyped and examined for antiretroviral resistance mutations. Data on gender of patient harboring the viruses and past history of antiretroviral treatment were also collected. RESULTS: Most viruses were either subtype B (61.44 percent) or subtype C (20.57 percent). Subtype C and F were more frequent in women (p < 0.00). The prevalence of subtypes was similar over the years studied. The most frequent RT mutations in all subtypes were M184V and mutations at codons 215, 41, 103, 67, 219, and 190. Mutations 41L, 210W, 215YF, and 74V were significantly more prevalent on subtype B, and the mutation 106M was significantly more prevalent on subtype C. The most frequent major PI mutations in all subtypes occurred at codons 46, 82, and 90. PR mutations 32I, 46I, and 84V were significantly more prevalent on subtype B. The minor PI mutations on codons 36, 93, and 63 were more prevalent on subtypes F, C, and B, respectively. CONCLUSION: We concluded that the predominant strain of HIV-1 in Paraná is subtype B, followed by subtype C. Some mutations at PR and TR had subtype predominance in accordance with other authors' report.
Subject(s)
Adult , Female , Humans , Male , Drug Resistance, Viral/genetics , Genetic Variation/genetics , HIV Infections/virology , HIV Protease/genetics , HIV Reverse Transcriptase/genetics , HIV-1 , Mutation/genetics , Antiretroviral Therapy, Highly Active , Cross-Sectional Studies , Genotype , HIV Infections/drug therapy , HIV-1 , Polymerase Chain Reaction , RNA, Viral/geneticsABSTRACT
O intuito de tal estudo é observar a prevalência de alteraçöes oculares numa populaçäo de pacientes portadores de HIV e avaliar a efetividade do exame oftalmoscópico sob midríase realizado por estudantes de medicina na detecçäo do envolvimento fundoscópico nestes pacientes e observar a ocorrência de sintomas oculares e alteraçöes fundoscópicas nestes pacientes. Foram avaliados pacientes portadores do HIV, em tratamento no Sistema Unico de Saúde em Curitiba, entre julho de 1996 e abril de 1997. As variáveis observadas foram: idade, sexo, presença de atividade laboral, existencia de sintomas oculares e as medicaçöes anti-retrovirais sistêmicas utilizadas. Um total de 120 pacientes foram submetidos a oftalmoscopia monocular direta por näo especialistas que, ao considerarem os pacientes como tendo alteraçöes fundoscópicas, encaminhavam-nos ao oftalmologista para realizaçäo de oftalmoscopia binocular indireta. Correlacionando-se o exame fundoscópico näo especializado, os sintomas e a avaliaçäo especializada dos pacientes, pode-se afirmar que o clínico pode desempenhar um papel importante no diagnóstico precoce de lesöes córioretinianas na Aids. Concluiu-se que os sintomas oculares mais frequentes foram visäo embaçada e moscas volantes e as alteraçöes oculares mais comuns, manchas algodonosas e turvaçäo de meios.(au)