H2S is synthesized via CBS/CSE pathway and triggered by cold conditions during Agaricus bisporus storage

BACKGROUND: H2S is proved to be functioning as a signaling molecule in an array of physiological processes in the plant and animal kingdom. However, the H2S synthesis pathway and the responses to cold conditions remain unclear in postharvest mushroom. RESULTS: The biosynthesis of H2S in the Agaricus bisporus mushroom tissues exhibited an increasing tendency during postharvest storage and was significantly triggered by cold treatment. The cystathionine clyase (AbCSE) and cystathionine b-synthase (AbCBS) genes were cloned and proved responsible for H2S biosynthesis. Furthermore, transcriptional and posttranscriptional regulation of AbCSE and AbCBS were crucial for the enzyme activities and subsequent H2S levels. However, the AbMST was not involved in this process. Moreover, the AbCSE and AbCBS genes displayed low identity to the characterized genes, but typical catalytic domains, activity sites, subunit interface sites, and cofactor binding sites were conserved in the respective protein sequences, as revealed by molecular modeling and docking study. The potential transcription factors responsible for the H2S biosynthesis in cold conditions were also provided. CONCLUSIONS: The H2S biosynthetic pathway in postharvest mushroom was unique and distinct to that of other horticultural products.

CBS promoter hypermethylation increases the risk of hypertension and stroke

OBJECTIVES: Cystathionine β-synthase is a major enzyme in the metabolism of plasma homocysteine. Hyperhomocysteinemia is positively associated with hypertension and stroke. The present study was performed to examine the possible effects of Cystathionine β-synthase promoter methylation on the development of hypertension and stroke. METHODS: Using quantitative methylation-specific PCR, we determined the Cystathionine β-synthase methylation levels in 218 healthy individuals and 132 and 243 age- and gender-matched stroke and hypertensive patients, respectively. The relative changes in Cystathionine β-synthase promoter methylation were analyzed using the 2-ΔΔCt method. The percent of the methylated reference of Cystathionine β-synthase was used to represent the Cystathionine β-synthase promoter methylation levels. RESULTS: In this study, the Cystathionine β-synthase promoter methylation levels of hypertensive and stroke participants were both higher than that of the healthy individuals (median percentages of the methylated reference were 50.61%, 38.05% and 30.53%, respectively, all p<0.001). Multivariable analysis showed that Cystathionine β-synthase promoter hypermethylation increased the risk of hypertension [odds ratio, OR (95% confidence interval, CI)=1.035 (1.025-1.045)] and stroke [OR (95% CI)=1.015 (1.003-1.028)]. The area under the curve of Cystathionine β-synthase promoter methylation was 0.844 (95% CI: 0.796-0.892) in male patients with hypertension and 0.722 (95% CI: 0.653-0.799) in male patients with stroke. CONCLUSION: Cystathionine β-synthase promoter hypermethylation increases the risk of hypertension and stroke, especially in male patients.

Diagnosis and Management of Classica Homocystinuria in Brazil: A Summary of 72 Late-Diagnosed Patients

Abstract This study described a broad clinical characterization of classical homocystinuria (HCU) in Brazil. This was a cross-sectional, observational study including clinical and biochemical data from 72 patients (60 families) from Brazil (South, n = 13; Southeast, n = 37; Northeast, n = 8; North, n = 1; and Midwest, n = 1). Parental consanguinity was reported in 42% of families. Ocular manifestations were the earliest detected symptom (53% of cases), the main reason for diagnostic suspicion (63% of cases), and the most prevalent manifestation at diagnosis (67% of cases). Pyridoxine responsiveness was observed in 14% of patients. Only 22% of nonresponsive patients on treatment had total homocysteine levels <100 mmol/L. Most commonly used treatment strategies were pyridoxine (93% of patients), folic acid (90%), betaine (74%), vitamin B12 (27%), and low-methionine diet + metabolic formula (17%). Most patients diagnosed with HCU in Brazil are late diagnosed, express a severe phenotype, and poor metabolic control. Milder forms of HCU are likely underrepresented due to underdiagnosis.

Effect of H2 S system on space learning and memory ability in PTSD model rat and on the frequency of spontaneous discharges of hippocampus neurons / 中国药理学通报

Aim To explore the behavior and CBS /H2 S levels of hippocampus in post traumatic stress dis-order (PTSD)rats and study the effect of exogenous H2 S on PTSD rats.Methods Single prolonged stress paradigm was adopted to prepare PTSD animal model. Morris water maze test was adopted to test space learn-ing and memory ability.CBS /H2 S content in hippo-campus tissue sample was measured using Western blot and methylene blue method.In vivo extracellular single unit recording was used to examine the frequency of spontaneous discharges of hippocampus neurons.Re-sults ① Escape latency increased and quadrant time (%)and platform crossing times decreased in Morris water maze test of PTSD group compared with normal group(P <0.01 ).CBS /H2 S level in hippocampus tis-sue of PTSD group also decreased compared with nor-mal group (P <0.01 ,P <0.05 ).② Escape latency decreased and quadrant time(%)and platform cross-ing times increased in Morris water maze test of NaHS+PTSD group compared with PTSD group(P <0.01 ).③ L-cysteine increased the frequency of spontaneous discharges of hippocampus neurons(P < 0.01 ).Con-clusions Learning and memory ability decrease in PTSD model rats owing to the inhibition of CBS /H2 S content in hippocampus tissue.The mechanism of be-havior improvement of H2 S on PTSD model rat is possi-bly related to the excitation of H2 S on frequency of spontaneous discharges of hippocampus neurons.

Relationship between Hcy and CBS-T833C gene polymorphism and breast cancer in Han of Xinjiang Population / 实用医学杂志

Objective To investigate the correlation between T833C gene polymorphism of cystathionine-βsynthase (CBS) and breast cancer in Xinjiang Han nationality. Methods CBS-T833C gene polymorphism of 200 breast cancer patients in Xinjiang Han nationality (breast cancer group) and 200 health examination people (control group) was analyzed by amplification refractory mutation system (ARMS) assay, Hcy levels and biochemical parameters were detected by automatic biochemical analyzer , the correlation between CBS-T833C gene polymorphism and breast cancer in Xinjiang Han nationality was analyzed. Results 1. Genotype TT,TC,CC of CBS T833C gene in the breast cancer group and the control group were respectively 55.5%, 32.0%, 12.5%and 53.0%, 34.0%, 12.0%. The G, A allele frequency of CBS T833C gen in the two groups were 71.5%,28.5%and 70.0%, 29.0%,the overall frequency distribution between the two groups was not statistically different among the three genotypes (P > 0.05). 2. Hcy levels in plasma of TC, CC genotype were higher than TT genotype in breast cancer group (P <0.05). Conclusion Increase of homocysteine levels may correlates with breast cancer in Xinjiang Han nationality, CBS-T833C gene polymorphism has no correlation with breast cancer in Xinjiang Han nationality.

Study on the relationship of metabolizing enzyme CBS G919A gene polymorphism with essential hypertension on Hui nationality in Xinjiang / 实用医学杂志

Objective To discuss the relationship of metabolizing enzyme Cystathionine βsynthase (CBS) G919A gene polymorphism with essential hypertension on Hui nationality in Xinjiang. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the gene polymorphism, blood lipid, blood sugar and other biochemical indicators were tested at the same time. Result The distribution frequency of genotype GG, GA, AA and A,G, of CBS G919A was 76.4%, 19.0%, 4.5%, which was of no significant difference compared with the control group (P > 0.05). Conclusion Gene polymorphism of CBS G919A was existed, which was no relationship with essential hypertension on Hui nationality in Xinjiang.

An improved preparing process for rivastigmine / 中国药科大学学报

@#An improved manufacturing process for rivastigmine(1)was developed by performing the condensation reaction of m-hydroxyacetophenone(4)with N-ethyl-N-methyl carbamoyl chloride, then Corey-Bakshi-Shibata(CBS)chiral reduction to(R)-3-(1-hydroxyethyl)phenyl ethyl(methyl)carbamate(2)and then mesylation with methanesulfonyl chloride and nucleophilic substitution with dimethylamine, respectively. To be successful, a crucial reductive process in the conversion of ketone(3)to chiralalcohol(2)had to be correctly understood and optimized via orthogonal experiment. The whole improved process was convenient for operation and purification, with completion of the synthesis of rivastigmine and an overall yield of 88%.

Genetic variation in genes involved in folate and drug metabolism in a south Indian population.

BACKGROUND: Genetic variations represented as single nucleotide polymorphisms (SNPs) vary across the world population. This genetic polymorphism (such as SNPs) plays an important role in pharmacogenomics. SNPs that affects cellular metabolism, by altering the enzyme activity, have an important role in therapeutic outcome. Allele frequencies in number of clinically relevant SNPs within south Indian populations are not yet known. Hence, we genotyped randomly selected unrelated south Indian subjects from different locations of south India representing the heterogeneous ethnic background of the population. MATERIALS AND METHODS: Common variants of MTHFD1, TYMS, SHMT1, MTR, MTRR, CBS and SULT1A1 gene polymorphisms were screened from healthy unrelated south Indian volunteers. Genotypes were determined using RFLP analysis of polymerase chain reaction-amplified products and confirmed by DNA sequencing. Chi-square test was performed to test for deviation from the Hardy-Weinberg equilibrium for each locus. RESULTS: Gene allele frequency for several polymorphisms in our study differed significantly between the populations of other nations reported for several of the SNPs. These results demonstrate that the populations in different geographic regions may have widely varying genetic allele frequencies for clinically relevant SNPs. CONCLUSION: The present study reports, for the first time, the frequency distribution of MTHFD1, TYMS, SHMT1, MTR, MTRR, CBS and SULTIA1 gene polymorphisms in a south Indian population. Population-specific genetic polymorphism studies will help in practicing pharmacogenomic principles in the clinics.

Análisis descriptivo de los niveles de homocisteína, polimorfismos C699T, C1080T y 844Ins68pb de la CBS, C677T de la MTHFR y otros factores de riesgo cardiovascular en una población colombiana con síndrome coronario agudo / Descriptive analysis of homocysteine levels, polymorphisms C699T, C1080T and 844Ins68pb of CBS, C677T from the MTHFR, and other cardiovascular risk factors in a Colombian population with acute coronary syndrome

La enfermedad cardiovascular es la primera causa de muerte en el mundo occidental. Por esta razón, es necesario describir los factores de riesgo conocidos, al igual que los factores genéticos, nutricionales y ambientales emergentes, como la hiperhomocisteinemia y la deficiencia de la vitamina B12 y de ácido fólico en la población colombiana, que permitan proponer estrategias comunitarias de control de la enfermedad. El objetivo de este estudio fue describir los factores de riesgo conocidos y los emergentes,principalmente la hiperhomocisteinemia y los polimorfismos relacionados con ella en pacientes con síndrome coronario agudo. El estudio incluyó 156 pacientes, a quienes se les cuantificó perfil lipídico, glucosa, creatinina, urea, homocisteína, vitamina B12 y ácido fólico, y se describieron las frecuencias de las variantes polimórficas c.677C/T, de la MTHFR (5,10-methylenetetrahydrofolate reductase y c.699C/T, c.1080 C/T y c.844ins68pb de CBS (Cystathionine â-Synthase). El 43,6% de los pacientes con síndrome coronario agudo correspondió a las mujeres y el 56,4% a los hombres. Los valores medios de colesterol total, cLDL, cHDL, glucosa, homocisteína, vitaminas B12 y ácido fólico, se encontraron en el rango normal. Sin embargo, se pudo observar que la homociste ína presentaba una tendencia al aumento con la edad, tanto en hombres como en mujeres. Los niveles de cHDL en el grupo de hombres y de mujeres, presentaron diferencia significativa (p=0,0379) e, igualmente, la diferencia fue significativa en los niveles de creatinina (p=0007), de vitamina B12 (p=0,0341) y en la diabetes mellitus (p=0,0436). Con este estudio se realizó una aproximación a la descripción de los niveles del perfil lipídico, glucemia, hiperhomocisteinemia y de polimorfismos en genes involucrados en la vía de la homocisteínametionina, en pacientes con enfermedad cardiovascular en la población colombiana...

Cardiovascular diseases are the main cause of death in the western world. Therefore, it is necessary to describe the associated genetic, nutritional and environmental risk factors, including hyperhomocysteinemia, vitamin B12 and folic acid deficiencies in the Colombian population. Through this survey we want to propose strategies to the community in order to control cardiovascular diseases. The goal of this study was to describe the known risk factors and also the emerging ones such as hyperhomocysteinemia and some polymorphisms, in a Colombian population Our study included 156 patients with acute coronary artery syndrome, whose lipid, glucose, creatinine, homocysteine, vitamin B12 and folic acid levels were measured and the identification of polymorphisms 677C/T, from the MTHFR and 699C/T, 1080C/T, 844ins68pb of CBS. Overall, 43.6% of patients with acute coronary artery syndrome corresponded to women, and 56.4% to men who participated in this study. The results of cholesterol CLDL, CHDL, glucose, homocysteine, vitamin B12 and folic acid levels were found in normal ranges. However, we were able to observe that the homocysteine presented a tendency to increase with age in men and women,the CHDL levels within the group of men and women showed a significant difference (p=0.0379)as well as in the levels of creatinine (p=0.0007) of vitamin B12 (p=0.0341) and diabetes mellitus (p=0.0436). In this study, we propose a rough description of the lipid, glycemia, and hyperhomocysteinemia levels and polymorphisms in genes involved the homocysteine-methionine metabolism in patients with cardiovascular disease in the population of Colombia...

Molecular Characterization and Functional Prediction of a Novel Leaf SAG Encoding a CBS-domain-containing Protein from Coleus blumei / 中国生物化学与分子生物学报

Leaf senescence is considered as one of important factors to decrease ornamental values of foliage plants. In the attempt to study and understand the molecular mechanism of leaf senescence, a senescent leaf cDNA library of Coleus blumei was constructed and a small EST library was obtained. According to the sequence of an EST fragment with a cystathionine beta synthase (CBS) domain, a novel leaf senescenceassociated gene (SAG) full-length cDNA encoding a CBS-domain-containing protein, denoted Cbcbs, was rapidly cloned using a strategy of RACE combined with cDNA library. The full length of the Cbcbs gene was 859 bp long (accession No. EF076754) and contained a 609 bp open reading frame (ORF) encoding a 202amino acid protein. One stop codon (TAA) was found in 5' UTR and one possible polyadenylation signal,AATAAA, and a pentanucleotide motif, ATTTA, were found in 3' UTR. The CbCBS contained a predicted mitochondrial targeting peptide in the N-terminal region, two conserved and intact CBS domains, four casein kinase Ⅱ (CK Ⅱ) phosphorylation sites, three protein kinase c (PKC) phosphorylation sites and one tyrosine sulfation (TS) site. Sequence comparisons and phylogenetic analysis showed that CbCBS was a novel senescence or stress-associated protein. The prediction analysis of secondary structure and three dimensional structure of CbCBS suggested that the chief function of the protein was decided by the CBS domain pair. The expression pattern of Cbcbs in leaves was analyzed by RT-PCR. It was demonstrated that Cbcbs gene was a senescence-associated gene (SAG) and expressed in all leaf stages, young stage (Y) being the lowest and terminal senescence stage (S3) being the highest, and was upregulated along with the leaf senescence.Function analysis showed that the mature CbCBS maybe acts as a sensor of cellular energy status and directly or indirectly regulates cellular energy levels to increase ATP content in mitochondria during periods of metabolic stress of senescent leaves.

Application of CBS in the integrated Chinese-western internal medicine teaching / 中华医学教育探索杂志

CBS(Case Based Study)teaching method is given in a case requiring students to answer a series of questions surrounding the case.In the teaching process,students of small groups are required to find their own solutions.Compared with tradition method,this mothed can improve students'academic performance(P

Processo de validação da escala de comportamento do treinador - versão atleta (ECT-A) / Validation process of the coach behavior scale - athlete version (ECT-A)

O objetivo deste estudo foi adaptar culturalmente e validar o questionário Coach Behavior Scale for Sport (CBS-S), de CÔTÉ et al. (1999) na Língua Portuguesa, denominado de Escala de Comportamento do Treinador - Versão Atleta (ECT-A). Inicialmente o ECT-A constou de 13 questões demográficas e 48 questões distribuídas em seis dimensões: Treinamento Físico, Treinamento Técnico, Preparação Mental, Estabelecimento de Objetivos, Reforço Pessoal Positivo e Reforço Pessoal Negativo. No Estudo 1 (80 atletas <47 homens e 33 mulheres>), os resultados da análise fatorial mostraram que a maioria das questões agrupou-se em dois fatores, contudo, a correlação entre algumas questões foi baixa, indicando a necessidade de revisões semânticas. No Estudo 2 (168 atletas <118 homens e 50 mulheres>), houve uma melhora nos resultados da análise fatorial, pois a dimensão de Reforço Pessoal Positivo apresentou um perfil similar ao CBS-S, mas a correlação entre oito questões foi baixa, levando à exclusão das mesmas. No Estudo 3 (222 atletas <91 homens e 131 mulheres>), a análise fatorial obteve, na maioria das questões, um perfil similar ao instrumento original (CBS-S) e a correlação entre as questões foi satisfatória (0,30). Os valores do Alpha de Cronbach, nas seis dimensões, foram 0,70 nos três estudos (N = 470), indicando uma fidedignidade satisfatória. Esses resultados foram indicativos para a validação do ECT-A, que na versão final ficou composto por 40 questões.

The purpose of the present study was to adapt and validate the version of the questionnaire Coach BehaviorScale for Sport -CBS-S- (CÔTÉ et al., 1999) into the Portuguese language, called Escala de Comportamento doTreinador - Versão Atleta (ECT-A). The ECT-A was composed by 13 demographic questions and 48 questionsdistributed into six dimensions: Physical Training, Technical Training, Mental Preparation, Goal Setting, PersonalPositive Reinforcement and Personal Negative Reinforcement. Results of Alpha Cronbach, in the six dimensions,were ³ 0,70, of the ECT-A in three studies (N = 470), indicating acceptable levels. In the Study 1, (80 athletes<47 men and 33 women>) results of factor analysis showed a grouped of majority of questions in two factorshowever correlations between some questions were low suggesting semantic revisions. In the Study 2, (168athletes <118 men and 50 women>) there was an improvement in the factor analysis, since Personal Positive Reinforcement dimension had a similar profile of the CBS-S, but correlation between 8 questions continuedlow and so they were excluded. In the Study 3, (222 athletes <91 men and 131 women>) factor analysis ofthe majority of the questions obtained a similar profile of the original instrument (CBS-S) and correlationsbetween questions were satisfactory (³ 0,30). These results showed evidences to validate the ECT-A ,whichfinal version was composed by 40 questions.

The Application of CBS in the Teaching of Medical Microbiology / 微生物学通报

CBS(Case Based Study)is a new teaching method,which is based on the case given.It requires students to answer questions according to the case. In the teaching,the students who are divided into groups,seek for resolvent by themselves. They should combine the information provided in the case,find the key answer and the relationship.The students gain the knowledge by reviewing and studying in the textbook,then analyse and solve questions with them. The application of CBS could make knowledge more relatively and systematic, moreover,it also make study more actively and initiatively.