SUMMARYTea has been considered a medicine and a healthy beverage since ancient times, but recently it has received a great deal of attention because of its antioxidant properties. Green tea polyphenols have demonstrated to be an effective chemopreventive agent. Recently, investigators have found that EGCG, one of the green tea catechins, could have anti-HIV effects when bound to CD4 receptor. Many factors can constitute important influences on the composition of tea, such as species, season, age of the leaf, climate, and horticultural practices (soil, water, minerals, fertilizers). This paper presents an HPLC analytical methodology development, using column RP-18 and mobile phase composed by water, acetonitrile, methanol, ethyl acetate, glacial acetic acid (89:6:1:3:1 v/v/v/v/v) for simultaneous determination and quantification of caffeine (CAF), catechin (C), epicatechin (EC) and epigallocatechin gallate (EGCG) in samples of Camellia sinensis (green tea) grown in Brazil and harvested in spring, in summer and in autumn, in comparison to Brazilian black tea, to samples of Japanese and Chinese green tea and to two standardized dry extracts of green tea. The method has been statistically evaluated and has proved to be adequate to qualitative and quantitative determination of the samples. Keywords: HPLC; EGCG; catechin; Camellia sinensis; Brazilian green tea. RESUMOMÉTODO RÁPIDO PARA DETERMINAÇÃO DE GALATO DE EPIGALOCATEQUINA (EGCG), EPICATEQUINA (EC), CATEQUINA (C) E CAFEÍNA (CAF) EM CHÁ VERDE USANDO CLAE. O chá vem sendo considerado uma bebida saudável e com propriedades medicinais desde tempos bem remotos, mas recentemente tem ganhado grande interesse no meio científico devido a suas atividades como antioxidante. Os polifenóis do chá verde vêm demonstrando possuir atividades quimiopreventivas. Recentemente, pesquisadores descobriram que o EGCG, a principal catequina do chá verde, poderia ter efeito anti-HIV quando acoplado ao receptor CD4. Muitos fatores podem influenciar de forma significativa na composição do chá, como sua espécie, estação que foi colhida, idade da folha, clima e técnicas de cultura (solo, irrigação e fertilizantes). Este artigo vem apresentar uma metodologia analítica desenvolvida para análise em CLAE, usando coluna de fase reversa C-18 e fase móvel composta de água, acetonitrila, metanol, acetato de etila, ácido acético glacial (89:6:1:3:1 v/v/v/v/v) para análise simultânea de cafeína (CAF), catequina (C), epicatequina (EC) e galato de epicatequina (EGCG) em amostras de chá verde, cultivado no Brasil, e colhidas nas estações de primavera, verão e outono; comparando-os com o chá preto brasileiro, amostras de chá verde japonês e chinês e também com dois extratos padronizados importados de chá verde. O método foi avaliado estatisticamente e se mostrou adequado para analisar qualitativamente e quantitativamente as amostras. Palavras-chave: CLAE; EGCG; catequina; Camellia sinensis; chá verde brasileiro.have found that EGCG, a green tea catechin, could have anti-HIV effects when bound to CD4 recept...

The study findings provide evidence of the clinical impact of the interaction between pharmacogenetic variability and environmental factors on ADRs induced by anti-TB drug therapy. Predictive pharmacogenetic testing and a comprehensive clinical history would therefore be helpful for identification and careful monitoring of patients at high risk of this complication.

Freeze-dried extracts from Camellia sinensis var. assamica IAC-259 cultivar named Brazilian green tea were prepared by hot water and ultrasound-assisted extractions using leaves harvested in spring and summer. Their caffeine and catechin contents were measured by high performance liquid chromatography-diode array detector. The antioxidant activity of the major green tea compounds and Brazilian green tea extracts was evaluated using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The levels of caffeine were higher in the summer samples (p < 0.05); otherwise, there were no significant variations related to the catechin contents between spring and summer samples. The sonication method using water/acetone as solvent had a high efficiency to extract not only epigallocatechin gallate but also epicatechin gallate (p < 0.05). Antioxidant activities of the Brazilian green tea extracts were not significantly different among seasons and extraction systems. The antioxidant data (IC50) of the Brazilian green tea extracts showed a significant correlation with their epigallocatechin gallate and epicatechin gallate contents (p < 0.05).

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Reverse phase-solid phase extraction from Cassia alata leaves (CaRP) was used to obtain a refined extract. Higher than wild-type sensitivity to CaRP was exhibited by 16 haploid Saccharomyces cerevisiae mutants with defects in DNA repair and membrane transport. CaRP had a strong DPPH free radical scavenging activity with an IC50 value of 2.27 μg mL−1 and showed no pro-oxidant activity in yeast. CaRP compounds were separated by HPLC and the three major components were shown to bind to DNA in vitro. The major HPLC peak was identified as kampferol-3-O-β-d-glucoside (astragalin), which showed high affinity to DNA as seen by HPLC-UV measurement after using centrifugal ultrafiltration of astragalin-DNA mixtures. Astragalin-DNA interaction was further studied by spectroscopic methods and its interaction with DNA was evaluated using solid-state FTIR. These and computational (in silico) docking studies revealed that astragalin-DNA binding occurs through interaction with G-C base pairs, possibly by intercalation stabilized by H-bond formation.

Plant extracts have a long history to be used in folk medicine. Cassia alata extracts are known to exert antibacterial activity but details on compounds and mechanism of action remain poorly explored. We purified and concentrated the aqueous leaf extract of C. alata by reverse phase-solid phase extraction and screened the resulting CaRP extract for antimicrobial activity. CaRP extract exhibited antimicrobial activity for Pseudomonas aeruginosa, Staphylococcus epidermidis, S. aureus, and Bacillus subtilis. CaRP also inhibited biofilm formation of S. epidermidis and P. aeruginosa. Several bacterial growth-inhibiting compounds were detected when CaRP extract was fractionated by TLC chromatography coupled to bioautography agar overlay technique. HPLC chromatography of CaRP extract yielded 20 subfractions that were tested by bioautography for antimicrobial activity against S. aureus and S. epidermidis. Five bioactive fractions were detected and chemically characterized, using high-resolution mass spectrometry (qTOF-MS/MS). Six compounds from four fractions could be characterized as kaempferol, kaempferol-O-diglucoside, kaempferol-O-glucoside, quercetin-O-glucoside, rhein, and danthron. In the Salmonella/microsome assay CaRP showed weak mutagenicity (MI < 3) only in strain TA98, pointing to a frameshift mutation activity. These results indicate that C. alata leaf extract contains a minimum of 7 compounds with antimicrobial activity and that these together or as single substance are active in preventing formation of bacterial biofilm, indicating potential for therapeutic applications.

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Alcohol dehydrogenases catalyse the reversible oxidation of alcohols to aldehydes or ketones, with concomitant reduction of NAD + or NADP + . Adh1p is responsible for the reduction of acetaldehyde to ethanol, while Adh2p catalyses the reverse reaction, the oxidation of ethanol to acetaldehyde. Lack of Adh1p shifts the cellular redox balance towards excess NADH/NADPH and acetaldehyde, while absence of Adh2p does the opposite. Yeast mutant adh1 had a slow growth rate, whereas adh2 grew like the isogenic wild-type (WT) during prediauxic shift fermentative metabolism. After 48 h WT and mutants reached the same number of viable cells. When exponentially growing (LOG) cells were exposed to calcofluor white, only mutant adh1 displayed an irregular deposition of chitin. Quantitative analyses of both LOG and stationaryphase cells showed that adh1 mutant contained significantly less ergosterol than cells of WT and adh2 mutant, whereas the erg3 mutant contained extremely low ergosterol pools. Both adh1 and adh2 mutants showed higher-than-WT resistance to heat shock and to H 2 O 2 but had WT resistance when exposed to ultraviolet (UV) light and the DNA cross-linking agent diepoxyoctane, indicating normal DNA repair capacity. Mutant adh1 was specifically sensitive to acetaldehyde and to membrane peroxidizing paraquat. Our results link the pleiotropic phenotype of adh1 mutants to low pools of ergosterol and to reductive stress, and introduce the two new phenotypes, resistance to heat shock and to H 2 O 2 , for the adh2 mutant, most probably related to increased ROS production in mitochondria, which leads to the induction of oxidative stress protection.