Highly selective alkylcoumarates and diterpenoids isolated from leaves of Baccharis quitensis: a search for new hit compounds against Trypanosoma cruzi.

In the present work, the hexane extract of aerial parts of Baccharis quitensis Kunth. was subjected to chromatographic fractionation to afford two alkyl phenylpropanoids: n-docosyl (E)-p-coumarate (1) and n-tetracosyl (E)-p-coumarate (2) as well as five diterpenes: ent-kaurenoic acid (3), grandifloric acid (4), 15ß-senecioyl-oxy-ent-kaur-16-en-19-oic acid (5), and 15-oxo-ent-kaurenoic acid (6). Using an ex-vivo assay with macrophages infected with Trypanosoma cruzi, compounds 1 and 2 demonstrated high potency against intracellular amastigotes, with EC50 values of 7.5 and 6.9 µM, respectively. Compound 6 revealed a moderate potency against T. cruzi, with an EC50 of 25.6 µM, and compounds 3-5 showed no effectiveness. Additionally, compounds 1-6 compounds presented no toxicity for mammalian cells to the highest tested concentration of 200 µM. Based on potency and the selectivity indexes of 1, 2 and 6, these compounds could be future candidates for optimisation studies for the design of prototypes against Chagas disease.

Highly selective alkylcoumarates and diterpenoids isolated from leaves of Baccharis quitensis: a search for new hit compounds against Trypanosoma cruzi

In the present work, the hexane extract of aerial parts of Baccharis quitensis Kunth. was subjected to chromatographic fractionation to afford two alkyl phenylpropanoids: n-docosyl (E)-p-coumarate (1) and n-tetracosyl (E)-p-coumarate (2) as well as five diterpenes: ent-kaurenoic acid (3), grandifloric acid (4), 15β-senecioyl-oxy-ent-kaur-16-en-19-oic acid (5), and 15-oxo-ent-kaurenoic acid (6). Using an ex-vivo assay with macrophages infected with Trypanosoma cruzi, compounds 1 and 2 demonstrated high potency against intracellular amastigotes, with EC50 values of 7.5 and 6.9 µM, respectively. Compound 6 revealed a moderate potency against T. cruzi, with an EC50 of 25.6 µM, and compounds 3–5 showed no effectiveness. Additionally, compounds 1–6 compounds presented no toxicity for mammalian cells to the highest tested concentration of 200 µM. Based on potency and the selectivity indexes of 1, 2 and 6, these compounds could be future candidates for optimisation studies for the design of prototypes against Chagas disease.

Polymorphisms of folate metabolism genes in patients with cirrhosis and hepatocellular carcinoma.

AIM: To evaluated the association of the risk factors and polymorphisms in MTHFR C677T, MTHFR A1298C, MTR A2756G and MTRR A66G genes. METHODS: Patients with cirrhosis (n = 116), hepatocellular carcinoma (HCC) (n = 71) and controls (n = 356) were included. Polymerase chain reaction followed by enzymatic digestion and allelic discrimination technique real-time PCR techniques were used for analysis. MINITAB-14.0 and SNPstats were utilized for statistical analysis. RESULTS: Showed that age ≥ 46 years (OR = 10.31; 95%CI: 5.66-18.76; P < 0.001) and smoking (OR = 0.47; 95%CI: 0.28-0.78; P = 0.003) were associated with cirrhosis. Age ≥ 46 years (OR = 16.36; 95%CI: 6.68-40.05; P < 0.001) and alcohol habit (OR = 2.01; 95%CI: 1.03-3.89; P = 0.039) were associated with HCC. MTHFR A1298C in codominant model (OR = 3.37; 95%CI: 1.52-7.50; P = 0.014), recessive model (OR = 3.04; 95%CI: 1.43-6.47; P = 0.0051) and additive model (OR = 1.71; 95%CI: 1.16-2.52; P = 0.0072) was associated with HCC, as well as MTR A2756G in the additive model (OR = 1.68; 95%CI: 1.01-2.77; P = 0.047), and MTRR A66G in the codominant model (OR = 3.26; 95%CI: 1.54-6.87; P < 0.001), dominant model (OR = 2.55; 95%CI: 1.24-5.25; P = 0.007) and overdominant model (OR = 3.05; 95%CI: 1.66-5.62; P < 0.001). MTR A2756G in the additive model (OR = 1.54; 95%CI: 1.02-2.33; P = 0.042) and smokers who presented at least one polymorphic allele for MTRR A66G (OR = 1.71; 95%CI: 0.77-3.82; P = 0.0051) showed increased risk for cirrhosis. There was no association between clinical parameters and polymorphisms. CONCLUSION: Age ≥ 46 years, alcohol habit and MTR A2756G, MTHFR A1298C and MTRR A66G polymorphisms are associated with an increased risk of HCC development; age ≥ 46 years, tobacco habit and the MTR A2756G polymorphism are associated with cirrhosis.

[Phenotypic characterization of yeasts isolated from the vaginal mucosa of adult women]. / Caracterização fenotípica de leveduras isoladas da mucosa vaginal em mulheres adultas.

PURPOSE: to characterize, phenotypically, yeasts isolated from the vaginal content of 223 symptomatic (S) and asymptomatic (A) adult women with vulvovaginitis, and to determine the clinical indicators which may lead to the appearance of signs and symptoms related to the mucosa involvement by this pathology. METHODS: a questionnaire with open and closed questions on epidemiological clinical data was applied initially. Then, mycological diagnosis with sowing in Chrom Agar Candida was done, followed by micro-morphological and biochemical identification. Specific methods for the detection of the virulence factors, proteinase and phospholipase were employed. Statistical analysis was performed through chi2 and Pearson's chi2 tests. RESULTS: the most prevalent species found was Candida albicans (87%, S and 67%, A) followed by Candida glabrata (4%, S e 17% A). The number of women reporting the use of contraceptives was higher among the symptomatic, 77%. In the two groups studied, about 87% of the women presented regular menstrual cycles and 57% were married with ages between 30 to 40 years old. Concerning the sexual practices, there has been concomitance among anal, oral and vaginal habits from the patients. Only Candida albicans produced the virulence factor phospholipase in 37.5% of them. Proteinase has been detected in Candida albicans, Candida glabrata and Candida parapsilosis. This latter virulence factor was mainly associated to isolates from symptomatic patients. CONCLUSIONS: it is a fact that the vaginal mucosa can be colonized and infected by yeasts, with several Candida species present. Nevertheless, Candida albicans is the most prevalent in the vaginal mucosa of adult women. It is evident the emergence of non-albicans Candida species, some of them with intrinsic resistance to azolics, such as Candida glabrata, Candida parapsilosis, Candida tropicalis, and Candida guillermondii, which can be explained by the inadequate use of medicines and empirical treatment.

Caracterização fenotípica de leveduras isoladas da mucosa vaginal em mulheres adultas / Phenotypic characterization of yeasts isolated from the vaginal mucosa of adult women

OBJETIVO: caracterizar fenotipicamente leveduras isoladas do conteúdo vaginal de 223 mulheres adultas, sintomáticas (S) e assintomáticas (A) para vulvovaginite, e determinar os indicadores clínicos que possivelmente levam ao surgimento de sinais e sintomas relacionados ao acometimento da mucosa por essa patologia. MÉTODOS: inicialmente foi aplicado um questionário, com questões abertas e fechadas, sobre dados clínicos epidemiológicos. Logo, ocorreu o diagnóstico micológico com semeadura em meio Chrom Agar Candida, identificação micromorfológica e bioquímica. Métodos específicos para detecção de fatores de virulência, proteinase e fosfolipase foram empregados. A análise estatística das variáveis foi estabelecida utilizando os testes χ2 e χ2 de Pearson. RESULTADOS: Candida albicans foi a espécie mais prevalente (87%, S e 67%, A), seguida de Candida glabrata (4%, S e 17%, A). O número de mulheres que referiram adoção de anticoncepcionais foi mais alto entre as sintomáticas, 77%. Nos dois grupos estudados, em torno de 87% apresentaram ciclos menstruais regulares, 57% das mulheres eram casadas com idade entre 30 a 40 anos. Em relação a práticas sexuais, houve para parte das pacientes, concomitância entre os hábitos, anal, oral e vaginal. Em relação à fosfolipase, apenas Candida albicans produziu este fator de virulência em 37,5%. A proteinase foi detectada em Candida albicans, Candida glabrata e Candida parapsilosis. Esse último fator de virulência esteve associado, principalmente, a isolados de pacientes sintomáticas. CONCLUSÕES: a colonização e infecção da mucosa vaginal por levedura é real com diversas espécies de Candida presentes. No entanto, Candida albicans se destaca como espécie prevalente em mucosa vaginal de mulheres adultas. Fica evidente a emergência de espécies de Candida não albicans, algumas com resistência intrínseca aos azólicos, tais como Candida glabrata, Candida...

PURPOSE: to characterize, phenotypically, yeasts isolated from the vaginal content of 223 symptomatic (S) and asymptomatic (A) adult women with vulvovaginitis, and to determine the clinical indicators which may lead to the appearance of signs and symptoms related to the mucosa involvement by this pathology. METHODS: a questionnaire with open and closed questions on epidemiological clinical data was applied initially. Then, mycological diagnosis with sowing in Chrom Agar Candida was done, followed by micro-morphological and biochemical identification. Specific methods for the detection of the virulence factors, proteinase and phospholipase were employed. Statistical analysis was performed through χ2 and Pearson’s χ2 tests. RESULTS: the most prevalent species found was Candida albicans (87%, S and 67%, A) followed by Candida glabrata (4%, S e 17% A). The number of women reporting the use of contraceptives was higher among the symptomatic, 77%. In the two groups studied, about 87% of the women presented regular menstrual cycles and 57% were married with ages between 30 to 40 years old. Concerning the sexual practices, there has been concomitance among anal, oral and vaginal habits from the patients. Only Candida albicans produced the virulence factor phospholipase in 37.5% of them. Proteinase has been detected in Candida albicans, Candida glabrata and Candida parapsilosis. This latter virulence factor was mainly associated to isolates from symptomatic patients. CONCLUSIONS: it is a fact that the vaginal mucosa can be colonized and infected by yeasts, with several Candida species present. Nevertheless, Candida albicans is the most prevalent in the vaginal mucosa of adult women. It is evident the emergence of non-albicans Candida species, some of them with intrinsic resistance to azolics, such as Candida glabrata, Candida parapsilosis, Candida tropicalis, and Candida guillermondii, which can be explained by the inadequate use of medicines and empirical treatment.