Note:

The glutathione-S-transferases (GSTs) (EC.2.5.1.18) are enzymes that participate in cellular detoxification of endogenous as well as foreign electrophilic compounds, function in the cellular detoxification systems and are evolved to protect cells against reactive oxygen metabolites by conjugating the reactive molecules to the nucleophile scavenging tripeptide glutathione (GSH, &amp / #61543 / -glu-cys-gly). The GSTs are found in all eukaryotes and prokaryotic systems, in the cytoplasm, on the microsomes, and in the mitochondria. Cytosolic GSTs have been grouped into seven distinct classes as: alpha (&amp / #61537 / ), mu (&amp / #61549 / ), pi (&amp / #61552 / ), sigma (&amp / #61555 / ), omega, theta (&amp / #61553 / ) and zeta (&amp / #61540 / ). In comparison with other GSTs, class theta enzymes have proven difficult to isolate and characterize. Two distinct theta GSTs have been identified in man, GSTT1-1 and GSTT2-2 three in the rat rGST1-1, rGSTT2-2 and 13-13 and one in the mouse. this study, a class theta GST (GSTT2-2), with high activity towards 1-MS was isolated and purified from bovine liver in 3% yield with a purification factor of 3-fold. The purification protocol included a sequential DEAE cellulose anion exchanger liquid chromatography column, S-hexylglutathione agarose affinity column, dye binding orange A and chromatofocusing columns. The enzyme activity and protein content decreased rapidly after the last step of purification. The purified GSTT2-2 showed significant activity only towards 1-MS as 77 nmole/min/mg. The GSTT2-2 purified from bovine liver had a molecular weigth (Mr) value of about 28,200 which was also confirmed by Western Blott Analysis. The purified farctions of GSTT2-2 with other kolon farctions were tested with anti GSTT2-2, antiGST alfa, antiGST mu and antiGST pi antibodies. The enzyme activities towards CDNB, 4-nitrobenzylchloride (NBC) and 1-menapthyl sulfate were measured as described by Habig and Jacoby.

ABSTRACT Equilibrium unfolding, single- and double-jump kinetic studies were conducted to determine the unfolding and refolding pathway of glutaredoxin 2. Structural changes for wild-type glutaredoxin 2 were monitored by far-ultraviolet circular dichroism and intrinsic tryptophan fluorescence for equilibrium unfolding and intrinsic tryptophan fluorescence for single- and double-jump kinetics studies. Glutaredoxin 2 possesses two tryptophan residues in domain 2. In order to monitor changes in domain 1, cysteine 9 at the active site cysteines, situated in domain 1, was labelled with an extrinsic fluorophore, AEDANS, and a mutant was created (Y58W glutaredoxin 2). The AEDANS labelled protein displayed decreased alpha-helical secondary structure and conformational stability. A high degree of cooperativity and similar conformational stability was observed during the two-state transition of the urea-induced equilibrium unfolding of both the wild-type and Y58W glutaredoxin 2 proteins therefore Y58W glutaredoxin 2 could be used to assess structural changes in the local environment of domain 1 during unfolding and refolding. Two phases of unfolding, the fast and slow phase, occurred for both the wild-type and Y58W proteins. The slow phase involves structural rearrangements that expose small amounts of surface area while the fast phase represents gross structural unfolding exposing large amounts of surface area. The isomerization of the Val48-Pro49 peptide bond to the trans conformation occurs during the slow phase and this isomerization is coupled to conformational unfolding of the protein. The structural separation of these phases could be represented by two structural units (unit x and unit y), these units do not represent domain 1 and 2. The units could also result in parallel refolding pathways with the folding of the x unit involving the fast and slow refolding phases and the folding of the y unit of structure is represented by the medium phase of refolding. The fast and slow phases are further separated as the fast phase represents the gross structural folding of glutaredoxin 2 for species with the Val48-Pro49 peptide bond in the native cis conformation. The development of the slow phase after extended unfolding delay periods during double-jump refolding studies, as well as the acceleration of the rate of the phase by the peptidyl prolyl isomerase hFKBP-12 proved that the phase involves a proline peptide bond iv isomerization. This phase represents a slow isomerization coupled with conformational folding similar to the slow unfolding phase. Complex unfolding and refolding kinetics indicated the involvement of kinetic intermediates during (un)folding.

Free radicals which have critical roles in living systems through their beneficial and detrimental effects play an important role in medical revolution in health. Radicals are produced in the cells and tissues of our bodies by various processes and reactions. Diabetes mellitus is an extremely common disease in the world which seems to be accompanied by a shortage of antioxidants and an increase in free radicals, the end result of oxidative stress. Glutathione S-Transferases (GST / EC 2.5.1.18) are found in enzymatic defense system which has a role in defending cells against potentially toxic and/or carcinogenic compounds. In this study, the changes in the activities and expressions of various GST isozymes in the liver of diabetic rats related to oxidative stress were studied. The effects of antioxidants, Vitamin C and &amp / #945 / -Lipoic acid on GST isozyme activities and mRNA expressions were also investigated. According to our results, diabetic rats exhibited decreased mRNA expressions of both GSTA2 and GSTM1 genes, but the activities of only GST Mu isozyme decreased in diabetic rats, compared to controls and GST Alpha isozyme activity remained unchanged in diabetic animals. Our results also showed that &amp / #945 / -Lipoic acid individually has no significant effect on both GSTA2 and GSTM1 gene expressions and activities. Furthermore, although the administration of Vitamin C alone showed no significant effect on all GST isozyme activities, it decreased GSTA2 mRNA expression significantly. The administration of Vitamin C and &amp / #945 / -Lipoic acid together affected both GSTA2 and GSTM1 mRNA expressions in control rats, but only GST Mu activity showed a significant change. The results of this study showed that, the administration of two antioxidants, &amp / #945 / -Lipoic acid and Vitamin C alone and together did not reverse the results of diabetes at the level of both gene expression and activities of GST isozymes.

Plants can not escape from biotic and abiotic stress factors such as, extreme temperatures, high light intensity, drought, UV radiation, heavy metals, and pathogen attack. Plants have versatile defens systems against such stress conditions. In this study, the role of glutathione S-transferases (GSTs) in cold stress conditions were examined. Glutathione S-transferases are the enzymes that detoxify natural and exogenous toxic compounds by conjugation with glutathione. Glutathione, an endogenous tripeptide, is important as reducing agent, nucleophilic scavenger, and alleviate the chemical toxicity in the plants by the reaction of GSTs. Glutathione conjugates can be transported to the vacuoles or apoplast and are generally much less toxic than the parent compounds. In plants there are four distinct families of the soluble GSTs, namely Phi (F), Type I / Zeta (Z), Type II / Tau (U), Type III / Theta (T), Type IV. By contrast with the mammalian families of GST, relatively little is known about the plant GST families. Up to date, there is not any study on GST isolation and characterization from Turkish red pine, in this respect, this study well play a frontier role the future research dealing with this topic. In this study, some properties of Turkish red pine GST activity towards CDNB (1-chloro-2,4 dinitrobenzene) were examined. The average specific activity of Turkish red pine GST towards CDNB was found as 200&plusmn / 50 (Mean&plusmn / SE, n= 18) nmole/min/mg cytosolic protein. GSTs in cytosol prepared from Turkish red pine needles retained its activity without loss for four weeks at -80&amp / #61616 / C. The rate of conjugation reactions were linear up to 0.8mg of Turkish red pine cytosolic protein and 0.4 mg cytosolic protein was routinely used. The Turkish red pine GST showed its maximum activity at pH 8.0 in 25 mM phosphate buffer and 42 &amp / #730 / C. The measurements were carried out at room temperature (RT) of 25 &amp / #61616 / C. Turkish red pine GST seemed to be saturated at 1 mM CDNB and 1 mM GSH concentrations. The Vmax and Km values of Turkish red pine GST for CDNB was 416nmole/min/mg protein and 0,8 mM, respectively, and for GSH 106.4 nmole/min/mg protein and 0.10 mM, respectively. Turkish red pine cytosol was applied on DEAE-Sepharose fast flow column but almost no purification was achieved with respect GST activity. In order to examine the effects of cold stress on Turkish red pine GST activity, the GST activity was determined in 240 seedlings at &ndash / 3&amp / #61616 / , 0&amp / #61616 / and 13 &amp / #61616 / C environmental temperatures. It was observed that GST activity was the highest at -3&amp / #730 / C and the lowest at 13&amp / #730 / C in both cold resistant and sensitive families with the exception of Yaylaalan and &Ccedil / ameli.

Helicoverpa armigera is a polyphagous pest. Due to excessive use of insecticides, the field populations of H. armigera have become resistant to synthetic pyrethroids by one or combination of three mechanisms / reduced penetration through the cuticle, decreased nerve sensitivity and enhanced metabolism by the detoxification enzymes especially glutathione S-transferases. In this study, gut sections of H. armigera were obtained from Adana and Antalya field populations and susceptible populations from Israel. Each gut section was homogenized separately in 1.0 ml, 40 mM and pH 7.5 phosphate buffers. GST activity was determined using CDNB as substrate. Product formation linearly increased up to 29.5&micro / g proteins in 20mM, pH 7.5 phosphate buffers. Maximum reaction rate was reached at 30&amp / #9702 / C. The Vmax and Km values for GST towards CDNB and GSH were calculated with Lineweaver-Burk and Eadie-Scatchard plots as CDNB Vmax / 6.54&micro / mol/min/mg, 6.35&micro / mol/min/mg , Km / 0.29mM, 0.28mM ,respectively and as GSH Vmax / 6.42&micro / mol/min/mg, 6.65&micro / mol/min/mg, Km / 0.22mM, 0.23mM, respectively. Cytosolic GST activity of each individual from Adana, Antalya and susceptible populations were determined under optimized conditions. The mean of GST activity in Adana population (n=50) and Antalya population (n=50) were found 7.824&micro / mol/min/mg and 9.518&micro / mol/min/mg, respectively. The mean of GST activity in susceptible population (n=50) was determined as 3.272&micro / mol/min/mg. According to these results, GST activities of Adana and Antalya field populations&rsquo / showed statistically significant increase (p&lt / 0.05) than susceptible H. armigera populations with ANOVA method. In addition, Antalya population showed statistically increase (p&lt / 0.05) GST activity than Adana.

京都大学 / 0048 / 新制・課程博士 / 博士(エネルギー科学) / 甲第21885号 / エネ博第386号 / 新制||エネ||75(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 片平 正人, 教授 森井 孝, 教授 木下 正弘 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DGAM

Conselho Nacional de Desenvolvimento Científico e Tecnológico / O oxigênio é fundamental para os vertebrados. No entanto, variações dos níveis de oxigênio na água podem provocar estresse oxidante em peixes porque privação de oxigênio seguida de reoxigenação forma espécies reativas de oxigênio (ERO) em células. Níveis intracelulares de ERO aumentados favorecem que moléculas de proteínas, fosfolipídios e ácidos nucleicos sofram alterações, vindo a prejudicar muitas funções celulares. No Pantanal, habitat do pacu, o nível de oxigênio varia circadianamente na água das lagoas rasas que acabam isoladas dos rios na seca. O pacu evoluiu sob a pressão contínua da exposição aos efeitos prejudiciais das ERO causados pelos pulsos de inundação. A melatonina, uma indolamina produzida na glândula pineal, influencia os níveis de atividade de enzimas antioxidantes que reduzem ERO, além de ser capaz de doar elétrons ou captar radicais livres de forma não enzimática. Os níveis de melatonina no pacu são mais altos no verão e menores no inverno. Isoenzimas de glutationa S-transferases que conjugam o tripetídeo glutationa com o 4-hidroxinonenal, aldeído derivado da peroxidação de ácidos graxos por ERO, são importantes para evitar alteração funcional de proteínas por ligação do 4-hidroxinonenal à sua estrutura. Neste trabalho procuramos relação entre estresse oxidante, níveis de atividades de glutationa S-transferase e melatonina, para estabelecer se a melatonina ajudaria pacus a superar os efeitos deletérios das espécies reativas de oxigênio. Ensaiamos atividades de isoenzimas de glutationa S-transferases no citosol de fígado de pacus mantidos em normoxia, hipoxia, reoxigenação e hiperoxia no inverno e no verão. Medimos o efeito da melatonina in vitro e in vivo sobre as atividades de isoenzimas de glutationa S-transferase. Medimos os efeitos do estresse oxidante sobre a ligação do 4-hidroxinonenal com proteínas nos fígados de pacus tratados com melatonina. Somente as isoenzimas que conjugam 4-hidroxinonenal com glutationa mostraram menor atividade no inverno em relação ao verão; outras isoenzimas de glutationa S-transferases não alteram suas atividades sazonalmente. In vitro a melatonina não alterou a atividade de isoenzimas de glutationa S-transferase que conjugam o 4-hidroxinonenal, mas inibiu outras isoenzimas de glutationa S-transferase. In vivo a melatonina aumentou a atividade encontrada no inverno das isoenzimas que conjugam o 4-hidroxinonenal para os níveis do verão. A ligação de 4-hidroxinonenal com proteínas foi menor em pacus inoculados com melatonina. Nossos resultados mostram que a melatonina pode influenciar os efeitos de ERO em fígado de pacus. Ficou claro que a melatonina do plasma mantém os níveis de atividade conjugadora de 4-hidroxinonenal do fígado em pacus e que a baixa produção de melatonina no inverno não é adequada para a conjugação do 4-hidroxinonenal em fígado de pacus. / Oxygen is vital for vertebrates. However, changes in the levels of dissolved oxygen in water might cause oxidative stress in fishes because the shortage of oxygen followed by reoxygenation originates reactive oxygen species (ROS) inside cells. Higher intracellular levels of ROS favor alterations of proteins, phospholipids and nucleic acid molecules, which result in impairment of many cell functions. In Pantanal, the pacus habitat, circadian variation of the oxygen levels occurs in water of the shallow lagoons that ended up isolated from the rivers along the dry season. Pacu has evolved under the pressure of continuous exposition to harmful effects of ROS caused by the annual inundation pulses. Melatonin, an indolamine produced by the pineal gland, influences the levels of activity of antioxidant enzymes that reduce ROS, and is capable of donating electrons or scavenge free radicals nonenzymatically. Pacus melatonin levels are higher during summer than in winter. Glutathione S-transferases isoenzymes that catalyze the conjugation of the tripeptide glutathione with 4-hydroxynonenal, an aldehyde derived from peroxidation of fat acids by ROS, are important to avoid functional alterations of proteins consequential to the binding of 4-hydroxynonenal to their structures. In this work, we searched for facts that linked oxidative stress, levels of activity of glutathione S-transferase and melatonin, in order to establish whether melatonin could help pacus to overcome the pernicious effects of reactive oxygen species. We carried out assays of glutathione S-transferases in liver cytosol of pacus kept under normoxia, hypoxia, reoxygenation and hyperoxia, in the summer and in the winter. We measured the effect of melatonin in vitro and in vivo on isoenzymes of glutathione S-transferases. We measured the effects of oxidative stress on the binding of 4-hydroxynonenal to proteins in liver of pacu treated with melatonin. Only isoenzymes that conjugate 4-hydroxynonenal with glutathione showed less activity during the winter in comparison to the summer; other isoenzymes did not have their activities changed seasonally. In vitro, melatonin did not change the activity of glutathione S-transferases isoenzymes that conjugate 4-hydroxynonenal, but inhibited other isoenzymes of glutathione S-transferase. In vivo, melatonin enhanced the liver activity of the glutathione S-transferase that conjugate 4-hydroxynonenal found in winter up to the levels found in summer. The binding of 4-hydroxynonenal to proteins was lower in liver cytosol from pacus injected with melatonin. Our findings show that melatonin can influence the effects of ROS in liver of pacu. It became evident that plasma melatonin maintains the liver levels of the conjugating activity of 4-hydroxynonenal and that the lower production of melatonin during winter is not adequate to the conjugation of 4-hydroxynonenal.

A doença inflamatória intestinal (DII) e a esclerose sistêmica (ES) são doenças inflamatórias crônicas de difícil diagnóstico e tratamento. A etiologia da DII e da ES ainda não é completamente compreendida, mas sabe-se que fatores genéticos, imunológicos e ambientais estão envolvidos na sua patogênese. A DII possui dois principais subtipos clínicos: a doença de Crohn (DC) e a retocolite ulcerativa (RCU), caracterizados pela inflamação do intestino delgado e/ou cólon. Evidências sugerem que o aumento do estresse oxidativo desempenha um papel importante na fisiopatologia da DII. A ES é uma doença inflamatória autoimune rara, caracterizada pela fibrose progressiva da pele e de órgãos internos. A hipótese de que o aumento do dano oxidativo pode iniciar o dano vascular e desencadear os eventos patológicos observados na ES vem sendo investigada. Genes e enzimas envolvidos na metabolização (Fase I) e detoxificação (Fase II) de xenobióticos são utilizados como marcadores de susceptibilidade para o desenvolvimento de doenças que possuem fatores ambientais como fatores de risco. Em uma reação de Fase I, as enzimas do Citocromo P450 (CYP) inserem um átomo de oxigênio em um substrato deixando-o eletrofílico e reativo, criando um sítio para posterior conjugação pelas enzimas de Fase II. As enzimas Glutationa S-tranferases (GST) de Fase II catalisam a conjugação da glutationa com uma grande variedade de compostos eletrofílicos, detoxificando substâncias endógenas e exógenas. A atividade catalítica aumentada das enzimas CYP, bem como a falha na detoxificação de metabólitos pelas GST pode contribuir para o aumento do estresse oxidativo. O objetivo deste estudo foi investigar o papel de polimorfismos nos genes que codificam enzimas de metabolização (CYP1A*2C e CYP2E1*5B) e detoxificação (GSTT1 nulo, GSTM1 nulo e GSTP1 Ile105Val) na susceptibilidade a estas doenças. O grupo de pacientes com DII era constituído por 235 indivíduos e o grupo controle por 241 indivíduos, todos eurodescendentes. Na ES, 122 pacientes (99 eurodescendentes e 23 afrodescendentes) e 329 controles (241 eurodescendentes e 87 afrodescendentes) foram analisados. Os polimorfismos CYP foram genotipados por PCR-RFLP, enquanto que os polimorfismos em GSTT1 e GSTM1 foram genotipados por PCR multiplex e PCR-RFLP para GSTP1. As frequências alélicas e genotípicas foram comparadas entre pacientes e controles usando o teste de Qui-Quadrado. A respeito dos resultados das análises em DII, as frequências alélicas e genotípicas dos polimorfismos CYP1A1*2C, CYP2E1*5B e GSTP1 Ile105Val, bem como as frequências genotípicas do polimorfismo de presença/ausência de GSTM1, foram similares nos três grupos de pacientes (DII, DC e RCU) quando comparados ao grupo controle (P>0,05). Observouse uma frequência significativamente aumentada do genótipo nulo de GSTT1 no grupo de pacientes com DII quando comparado ao grupo controle [0,28 vs 0,18; χ² com Yates P=0,02; OR=1,71 (IC 95% 1,09 –2,71)]. Quando separamos o grupo de pacientes em DC ou RCU, esta frequência permaneceu significativamente aumentada somente no grupo de pacientes com RCU comparado ao grupo controle [0,29 vs 0,18; χ² com Yates P=0,035; OR=1,84 (IC 95% 1,03 –3,24)]. Com relação aos resultados das análises na ES, uma frequência significativamente aumentada do genótipo *1A/*1A (P=0,03; 0,74 vs. 0,61) e do alelo *1A (P=0,013; 0,86 vs 0,78; OR=0,57, IC 95% 0,36–0,90) do polimorfismo CYP1A1*2C foi observada entre os indivíduos controles eurodescendentes. Em contrapartida, a frequência do alelo *2C estava significativamente aumentada entre os pacientes de mesma etnia (P=0,013; 0,22 vs 0,14; OR=1,75, IC 95% 1,11–2,74). Com relação às frequências alélicas e genotípicas dos polimorfismos CYP2E1*5B e GSTP1 Ile105Val, e as frequências genotípicas do polimorfismo de presença/ausência de GSTM1, nenhuma diferença significativa foi observada quando os grupos de pacientes de ambas as etnias foram comparados aos grupos controle (P>0,05). Uma frequência significativamente aumentada do genótipo nulo de GSTT1 [0,29 vs 0,18; χ² com Yates P=0,035; OR=1,85 (IC 95% 1,03–3,29)], bem como uma alta frequência da dupla deleção de GSTT1/GSTM1 [0,19 vs 0,08; χ² com Yates P=0,007; OR=2,62 (IC 95% 1,25 –5,46)], foi observada no grupo de pacientes comparado aos controles (eurodescendentes). Estas associações não se repetiram entre indivíduos afrodescendentes. Concluindo, nossos resultados sugerem que o genótipo nulo de GSTT1 está associado à susceptibilidade a DII e pode influenciar na definição do curso da doença para a RCU. Além disso, o genótipo nulo de GSTT1 sozinho ou em combinação com o genótipo nulo de GSTM1 é um fator genético de susceptibilidade para a ES, enquanto que o genótipo *1A/*1A ou a presença do alelo *1A do polimorfismo CYP1A1*2C pode exercer um papel protetor contra o desenvolvimento da ES em indivíduos eurodescendentes. / Inflammatory bowel disease (IBD) and systemic sclerosis (SSc) are chronic inflammatory diseases of difficult diagnosis and treatment. The etiology of IBD and SSc is not completely understood but it is known that genetic, immunologic and environmental factors are involved in its pathogenesis. Crohn’s disease (CD) and ulcerative colitis (UC) are the two major subtypes of IBD, characterized by inflammation of the small intestine and/or colon. Evidences suggest that the increase of oxidative stress plays an important role in the pathophysiology of IBD. SSc is a rare autoimmune inflammatory disease of the connective tissue characterized by progressive fibrosis of the skin and internal organs. The hypothesis that the increase of oxidative stress can initiate vascular damage and triggers the pathological events in SSc has been investigated. Genes and enzymes involved in metabolism (Phase I) and detoxification (Phase II) of xenobiotics are used as markers of susceptibility to the development of diseases that have environmental factors as risk factors. In a Phase I reactions, the Cytochrome P450 (CYP) enzymes insert an oxygen atom in a substrate that making it more electrophilic and reactive, and creating a site for subsequent conjugation by Phase II enzymes. Phase II Glutathione S-transferases (GSTs) enzymes catalyze the conjugation of glutathione with a variety of electrophilic compounds, detoxifying endogenous and exogenous substances. A higher catalytic activity of CYP enzymes, as well as the failure in detoxifying of metabolites by GST enzymes may to contribute for the increase of oxidative stress. The aim of this study was investigated the role of polymorphisms in genes coding Phase I enzymes (CYP1A*2C and CYP2E1*5B) and Phase II (GSTT1 null, GSTM1 null and GSTP1 Ile105Val) in susceptibility to these diseases. IBD group was constituted by 235 patients and the control group by 241 individuals, all European-derived. In SSc group, 122 patients (99 European-derived and 23 African-derived) and 329 controls (241 European-derived and 87 African-derived) were analyzed. The CYP polymorphisms were genotyped by PCR-RFLP, whereas polymorphisms in GSTM1 and GSTT1 were genotyped by multiplex PCR and PCRRFLP for GSTP1. Allelic and genotypic frequencies were compared between patients and controls using the Chi-square test. Concerning IBD, allelic and genotypic frequencies of CYP1A1*2C, CYP2E1*5B and GSTP1 Ile105Val polymorphisms, as well as genotypic frequencies of GSTM1 presence/absence polymorphism were similar in all groups patients (IBD, CD, and UC) and controls (P>0.05). We observed a significantly increased frequency of GSTT1 null genotype in IBD group as compared to controls [0.28 vs. 0.18, χ ² with Yates P=0.02, OR=1.71 (95% CI 1.09 – 2.71)]. When patients were classified in CD or UC group, this frequency remained significantly increased only among UC patients [0.29 vs. 0.18, χ ² with Yates P=0,035, OR=1.84 (95% CI 1.03 – 3.24)] as compared to controls. Regarding results in SSc, a frequency significantly increased of *1A/*1A genotype (P=0.03; 0.74 vs. 0.61) and *1A allele (P=0.013; 0.86 vs 0.78; OR=0.57, 95% CI 0.36–0.90) from CYP1A1*2C polymorphism was observed among European-derived controls. On the other hand, the frequency of *2C allele was significantly increased among patients of same ethnic group (P=0.013; 0.22 vs 0.14; OR=1.75, 95% CI 1.11–2.74). The allelic and genotypic frequencies of CYP2E1*5B and GSTP1 Ile105Val polymorphisms, as well as genotypic frequencies of GSTM1 presence/absence polymorphism were similar between SSc patients and controls of both ethnic groups (P>0.05). We observed a significantly increased frequency of GSTT1 null genotype [0.29 vs. 0.18, χ ² with Yates P=0.035, OR=1.85 (95% CI 1.03–3.29)], as well as an increased frequency of GSTT1/GSTM1 double-null in SSc patients as compared to controls [0.19 vs. 0.08; χ ² with Yates P=0.007, OR=2.62 (95% CI 1.25 – 5.46)]. These associations were exclusive to European-derived individuals. In conclusion, our results suggest that the GSTT1 null genotype is associated with susceptibility to IBD and may influence in defining the course of the disease for RCU. Furthermore, the GSTT1 null genotype alone or combined with GSTM1 null genotype is a susceptibility genetic factor to SSc, while the *1A/*1A genotype or the presence of *1A allele from CYP1A1*2C polymorphism may plays a protector role in SSc development in Brazilian Europeanderived individuals.

S.absconditiflora is one of the endemic Salvia species grown in Turkey, which is consumed as a herbal tea. Because of the presence of high amounts of vesicles on their leaves, S.absconditiflora is very rich in active compounds. S.absconditiflora water extract was investigated for its antioxidant capacity by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Total phenolic and total flavonoid contents were quantified by spectrophotometric methods. LC-MS/MS analyses revealed the presence and quantities of caffeic acid, luteolin rutin and coumaric acid. Cytotoxic effects of water extract of S.absconditiflora on breast cancer cell lines (MCF-7 and MDA-MB-231) were examined via XTT colorimetric assay and Trypan Dye Exclusion cell viability assay. IC50 values for each cell line at 24 and 48 hours were determined. The results indicated that water extract of leaves of S.absconditiflora could inhibit cell proliferation in MCF-7 and MDA-231 cells in dose dependent but not in time dependent manner. Effects of S.absconditiflora water extract on the expression of glutathione-S-transferases (GSTs) in MCF-7 and MDA-MB-231 cells were investigated with qRT-PCR technique. IC50 values calculated in XTT experiment for 24h incubation was used as cytotoxic extract concentration. It was found that treatment of MCF-7 cells with 1,558 mg/ml of extract enhanced an increase in expression as 2 and 2,8 fold in GSTO1 and GSTZ1 genes, respectively. Treatment of MDA-MB-231 cells with 1,131 mg/ml of extract resulted in 1,57 fold increase for GSTO1 and 1,56 fold increase for GSTZ1.