Vaccines against toxoplasma gondii : challenges and opportunities

Development of vaccines against Toxoplasma gondii infection in humans is of high priority, given the high burden of disease in some areas of the world like South America, and the lack of effective drugs with few adverse effects. Rodent models have been used in research on vaccines against T. gondii over the past decades. However, regardless of the vaccine construct, the vaccines have not been able to induce protective immunity when the organism is challenged with T. gondii, either directly or via a vector. Only a few live, attenuated T. gondii strains used for immunization have been able to confer protective immunity, which is measured by a lack of tissue cysts after challenge. Furthermore, challenge with low virulence strains, especially strains with genotype II, will probably be insufficient to provide protection against the more virulent T. gondii strains, such as those with genotypes I or II, or those genotypes from South America not belonging to genotype I, II or III. Future studies should use animal models besides rodents, and challenges should be performed with at least one genotype II T. gondii and one of the more virulent genotypes. Endpoints like maternal-foetal transmission and prevention of eye disease are important in addition to the traditional endpoint of survival or reduction in numbers of brain cysts after challenge

Pronounced between-subject and circadian variability in thymidylate synthase and dihydropyrimidine dehydrogenase enzyme activity in human volunteers

AIMS: The enzymatic activity of dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) are important for the tolerability and efficacy of the fluoropyrimidine drugs. In the present study, we explored between-subject variability (BSV) and circadian rhythmicity in DPD and TS activity in human volunteers. METHODS: The BSVs in DPD activity (n = 20) in peripheral blood mononuclear cells (PBMCs) and in plasma, measured by means of the dihydrouracil (DHU) and uracil (U) plasma levels and DHU : U ratio (n = 40), and TS activity in PBMCs (n = 19), were examined. Samples were collected every 4 h throughout 1 day for assessment of circadian rhythmicity in DPD and TS activity in PBMCs (n = 12) and DHU : U plasma ratios (n = 23). In addition, the effects of genetic polymorphisms and gene expression on DPD and TS activity were explored. RESULTS: Population mean (± standard deviation) DPD activity in PBMCs and DHU : U plasma ratio were 9.2 (±2.1) nmol mg(-1) h(-1) and 10.6 (±2.4), respectively. Individual TS activity in PBMCs ranged from 0.024 nmol mg(-1) h(-1) to 0.596 nmol mg(-1) h(-1) . Circadian rhythmicity was demonstrated for all phenotype markers. Between 00:30 h and 02:00 h, DPD activity in PBMCs peaked, while the DHU : U plasma ratio and TS activity in PBMCs showed trough activity. Peak-to-trough ratios for DPD and TS activity in PBMCs were 1.69 and 1.62, respectively. For the DHU : U plasma ratio, the peak-to-trough ratio was 1.43. CONCLUSIONS: BSV and circadian variability in DPD and TS activity were demonstrated. Circadian rhythmicity in DPD might be tissue dependent. The results suggested an influence of circadian rhythms on phenotype-guided fluoropyrimidine dosing and supported implications for chronotherapy with high-dose fluoropyrimidine administration during the night

Análise de variáveis fisiológicas de adolescentes com diagnóstico clínico de asma leve intermitente ou leve persistente quando submetidos a hipóxia aguda e teste de esforço máximo Analysis of physiological variables during acute hypoxia and maximal stress test in adolescents clinically diagnosed with mild intermittent or mild persistent asthma

OBJETIVO: Analisar variáveis fisiológicas de adolescentes com diagnóstico clínico de asma quando submetidos a teste de hipóxia aguda e de esforço máximo. MÉTODOS: Estudo descritivo transversal composto por 48 adolescentes (12-14 anos), divididos em três grupos: 12 no grupo asma leve intermitente (ALI), 12 no grupo asma leve persistente (ALP) e 24 no grupo controle. Todos foram submetidos a teste de hipóxia aguda e a teste de esforço máximo. Características antropométricas foram coletadas, e variáveis funcionais foram determinadas antes e após o teste de esforço máximo. Em condições de hipóxia aguda, foram registrados o tempo de descida e o tempo de recuperação de SpO2 durante repouso. RESULTADOS: Não foram encontradas diferenças significativas nas variáveis antropométricas nem nas variáveis ventilatórias durante o teste de esforço entre os grupos. Foram encontradas diferenças significativas na pressão de oxigênio com 50% de saturação da hemoglobina antes do teste e na PaO2 antes do teste entre os grupos ALP e controle (p = 0,0279 e p = 0,0116, respectivamente), assim como na tensão de extração de oxigênio antes do teste entre os grupos ALI e ALP (p = 0,0419). Não houve diferenças significativas nos tempos de SpO2 em quaisquer das condições estudadas. O consumo de oxigênio e a eficiência da respiração foram semelhantes entre os grupos. O uso de um broncodilatador não trouxe vantagens nos resultados no teste de hipóxia. Não foram encontradas correlações entre o teste de hipóxia e as variáveis fisiológicas. CONCLUSÕES: Nossos achados sugerem que os adolescentes com asma leve persistente têm uma melhor capacidade de adaptação à hipóxia comparado aos com outros tipos de asma.<br>OBJECTIVE: To analyze adolescents clinically diagnosed with asthma, in terms of the physiological changes occurring during acute hypoxia and during a maximal stress test. METHODS: This was a descriptive, cross-sectional study involving 48 adolescents (12-14 years of age) who were divided into three groups: mild intermittent asthma (MIA, n = 12); mild persistent asthma (MPA, n = 12); and control (n = 24). All subjects were induced to acute hypoxia and were submitted to maximal stress testing. Anthropometric data were collected, and functional variables were assessed before and after the maximal stress test. During acute hypoxia, the time to a decrease in SpO2 and the time to recovery of SpO2 (at rest) were determined. RESULTS: No significant differences were found among the groups regarding the anthropometric variables or regarding the ventilatory variables during the stress test. Significant differences were found in oxygen half-saturation pressure of hemoglobin prior to the test and in PaO2 prior to the test between the MPA and control groups (p = 0.0279 and p = 0.0116, respectively), as was in the oxygen extraction tension prior to the test between the MIA and MPA groups (p = 0.0419). There were no significant differences in terms of the SpO2 times under any of the conditions studied. Oxygen consumption and respiratory efficiency were similar among the groups. The use of a bronchodilator provided no significant benefit during the hypoxia test. No correlations were found between the hypoxia test results and the physiological variables. CONCLUSIONS: Our findings suggest that adolescents with mild persistent asthma have a greater capacity to adapt to hypoxia than do those with other types of asthma

<b>18</b>β<b>-glycyrrhetinic acid inhibits rotavirus replication in culture</b>

<p>Abstract</p> <p>Background</p> <p>Glycyrrhizin (GA) and primary metabolite 18β-glycyrrhetinic acid (GRA) are pharmacologically active components of the medicinal licorice root, and both have been shown to have antiviral and immunomodulatory properties. Although these properties are well established, the mechanisms of action are not completely understood. In this study, GA and GRA were tested for the ability to inhibit rotavirus replication in cell culture, toward a long term goal of discovering natural compounds that may complement existing vaccines.</p> <p>Methods</p> <p>Epithelial cells were treated with GA or GRA various times pre- or post-infection and virus yields were measured by immunofluorescent focus assay. Levels of viral proteins VP2, VP6, and NSP2 in GRA treated cells were measured by immunoblot to determine if there was an effect of GRA treatment on the accumulation of viral protein.</p> <p>Results</p> <p>GRA treatment reduced rotavirus yields by 99% when added to infected cultures post-- virus adsorption, whereas virus yields in GA treated cultures were similar to mock treated controls. Time of addition experiments indicated that GRA-mediated replication inhibition likely occurs at a step or steps subsequent to virus entry. The amounts of VP2, VP6 and NSP2 were substantially reduced when GRA was added to cultures up to two hours post-entry.</p> <p>Conclusions</p> <p>GRA, but not GA, has significant antiviral activity against rotavirus replication <it>in vitro</it>, and studies to determine whether GRA attenuates rotavirus replication <it>in vivo</it> are underway.</p