Resonant Recognition Model Study for Interactions between SARS CoV 2 and Human Proteins

This study was devoted to the Resonant Recognition Model (RRM) analysis of SARS-CoV-2 proteins and their possible interaction with other human proteins, specifically, SARS CoV replicases and methyl transferases, were tested, via RRM analysis, for possible interactions with host CD4 T receptor proteins and prohibitins which participate in human organism response to viral infections. The following protein sequences were studied: twenty human SARS coronavirus methyltransferase proteins, eight replicase proteins, twenty-one prohibitin proteins, and eleven CD4 -T-cell surface antigens T4 proteins. Results revealed RRM peaks at f1=0.07349 and f2=0.2839. The peak at f1 was also common for interaction between SARS-CoV-2 methyl transferases and human prohibitins, where opposite phase suggest binding between these proteins during viral infection. This interaction was not supported for viral methyltransferase and human CD4 receptors (72.4 o phase shift). Viral replicases exhibited opposite phase interaction with both prohibitins and CD4 receptors. Overall, RRM revealed common RRM frequencies for both replicases and methyl transferases, and added plausibility to interactions between SARSCoV2 methyl transferase and human prohibitin, as well as between SARS Cov2 replicase and human prohibitin and CD4 T-cell receptors(AU)

Este estudio se dedicó al análisis mediante el Modelo de Reconocimiento Resonante (RRM) de las proteínas del SARS-CoV-2 y su posible interacción con otras proteínas humanas, específicamente, fueron analizadas las replicasas de SARS CoV y las metiltransferasas, mediante análisis RRM, para detectar posibles interacciones con las Proteínas del receptor CD4 T y las prohibitinas humanas, las cuales participan en la respuesta del organismo humano a las infecciones virales. Se estudiaron las siguientes secuencias de proteínas: veinte proteínas metiltransferasas del coronavirus del SARS humano, ocho replicasas, veintiuna prohibitinas y once proteínas T4 de antígenos de superficie de células T CD4. Los resultados revelaron picos de RRM en f1 = 0.07349 y f2 = 0.2839. El pico en f1 también fue común para la interacción entre las metiltransferasas del SARS-CoV-2 y las prohibitinas humanas, donde la fase opuesta sugiere la unión entre estas proteínas durante la infección viral. Esta interacción no fue apoyada para la metiltransferasa viral y los receptores CD4 humanos (cambio de fase de 72,4 o). Las réplicas virales exhibieron una interacción de fase opuesta tanto con las prohibitinas como con los receptores CD4. En general, el análisis de RRM reveló frecuencias comunes de RRM para replicasas y metiltransferasas, y apoyó plausibilidad de las interacciones entre la metiltransferasa de SARSCoV2 y la prohibitina humana, así como entre la replicasa de Cov2 del SARS con la prohibitina humana y los receptores de células T CD4(AU)

Tratamiento de la enfermedad de Parkinson temprana / Treatment of early Parkinson's disease

RESUMEN La enfermedad de Parkinson (EP) es principalmente una enfermedad de pacientes ancianos. Es un trastorno multifacético que comprende síntomas motores y no motores en todas las etapas de la enfermedad. Esta revisión busca integrar los datos de las opciones de tratamiento más recientes con los datos de las terapias establecidas, a fin de proporcionar una referencia actualizada basada en la evidencia para los médicos que tratan la EP temprana, con medicamentos que puedan usarse como alternativa a la levodopa. El enfoque de los médicos para el tratamiento de la enfermedad de Parkinson (EP) temprana debe tener en cuenta numerosos aspectos, entre ellos, cómo informar al paciente sobre el diagnóstico y la decisión crítica de qué terapia adoptar y cuándo iniciarla. El tratamiento del trastorno motor asociado con la EP temprana debe considerar varios factores cruciales, como la edad de inicio, las comorbilidades y los requisitos funcionales del paciente, y no se puede resumir en una fórmula simple. En pacientes más jóvenes (es decir, antes de la edad de 70 años) y en aquellos sin altos requisitos funcionales, el tratamiento generalmente se inicia con agonistas de dopamina y / o inhibidores de la enzima monoaminooxidasa-B (MAO- B I). En pacientes más jóvenes, la levodopa se debe agregar a los agonistas de la dopamina y / o MAO-B I, según lo requiera la progresión de la enfermedad, mientras que en los pacientes mayores, cuando la respuesta a la levodopa sola no es satisfactoria, los agonistas de la dopamina o los inhibidores de la catecol-O- metiltransferasa pueden posteriormente ser agregados.

SUMMARY Parkinson's disease (PD) is primarily a disease of elderly patients. Is a multifaceted disorder comprised of both motor and non-motor symptoms at all stages of the disease. This review seeks to integrate data from the newest treatment options with data from established therapies, so as to provide an up-to- date evidence-based reference for clinicians treating early PD, with medications that can be used as an alternative to levodopa. The clinicians' approach to the treatment of early Parkinson's disease (PD) should take into account numerous aspects, including how to inform a patient upon diagnosis and the critical decision of what therapy to adopt and when to start it. The treatment of the motor disorder associated with early PD needs to consider several crucial factors, such as age at onset, comorbidities, and the patient's functional requirements, and cannot be summarized in a simple formula. In younger patients (i.e., before the age of 70) and in those without high functional requirements, treatment is usually initiated with dopamine agonists and/or monoamine oxidase-B enzyme inhibitors (MAO-B I). In younger patients, levodopa should be added to dopamine agonists and/or MAO-B I, as required by disease progression, whereas in older patients, when response to levodopa alone is not satisfactory, dopamine agonists or catechol-O- methyltransferase inhibitors may subsequently be added.

The association of catechol-O-methyl transferase gene polymorphism with the clinical efficacy of risperidone in the treatment and cognitive function in patients with schizophrenia / 中华行为医学与脑科学杂志

Objective To investigate the relationship between the catechol-O-methyl transferase (COMT) gene polymorphism and the clinical efficacy and cognitive function of risperidone in the treatment of schizophrenia.Methods 105 cases of Han Chinese patients with schizophrenia who were treated with risperidone for 12 weeks and healthy controls of 168 cases were collected.The effect of the drug therapy with the PANSS,digit vigilance test,Raven Standard Progressive Matrices,forward and backward subtests of the digit span test were evaluated,and then the rs 165599,rs4680,rs6267,rs737865 loci in COMT gene were detected.Results (1)rs737865 was not the polymorphic locus in this sample.(2) There was statistically significant between schizophrenia patients and controls in the distribution of allele frequency and genotype frequency in rs4680 (x2=8.16,P=0.02).Haplotype GA in rs165599-rs4680 was statistically significant in schizophrenia patients and controls (x2 =4.35,P =0.04).(3) After treatment,the total score ((47.64±5.75) points),subscale scores (positive symptoms (11.66±2.90) points,negative symptoms (13.79±3.18)points,general psychotic symptoms (22.09±3.59) points) and scores of five factors model in PANSS decreased and the difference was significant (P＜0.05);the scores of digital cancellation test increased significantly compared with those before treatment(t value respectively were 12.34,10.17,4.34,all P＜0.05);the scores of forward and backward subtests of the digit span test were significantly increased compared with those before treatment (t=-5.57,P=0.00) and Raven standard reasoning test scores had increased significant (t=-19.05,P=0.00).(4) The difference of instantaneous memory score changes among rs 165599 genotypes was statistically significant after treatment (F=4.06,P=0.02).(5) The difference of negative syndromes of PANSS among rs 165599 genotypes was statistically significant after treatment (F=3.11,P=0.049).(6) The difference of negative symptoms (F=4.64,P=0.01),cognitive impairment (F=3.21,P =0.045) and instantaneous memory (F=4.86,P=0.03) among rs 6267 genotype were statistically significant after treatment.Condusion Risperidone can effectively improve the psychotic symptoms of schizophrenia patients,and promote the recovery of cognitive function.Rs165599-rs4680 haplotype GA might be risk factor for the onset of schizophrenia.

Correlation of catechol-O-methyl transferase Vall58Met polymorphism with prepulse inhibition of the auditorystartle reflex in patients with schizophrenia / 中国心理卫生杂志

Objective:To investigate the association between catechol-O-methyl transferase (COMT)Vall58Met polymorphism and prepulse inhibition of the auditory startle reflex (PPI) in patients with schizophrenia.Methods:Totally 178 patients with schizophrenia and 190 healthy volunteers were recruited.The auditory startle reflex was detected by using SR-HLAB monitoring system.The indexed of the auditory startle reflex included the amplitude,habituation％ and PPI30,PPI60,PPI120 (the lead interval was set 30 ms,60 ms,120 ms).COMT Vall58Met polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP).The differences of PPI among COMT genotypes were compared.Results:Compared to the healthy volunteers,patients with schizophrenia had a significant lower the amplitude of auditory startle reflex[(563± 460) mV vs (695 ± 447) mY,P ＜ 0.05] and habituation％ [(32 ± 46) vs (48 ± 33),P ＜ 0.01] as well as the ％PPI120[(27 ± 5) vs (35 ± 3),P ＜ 0.05].The significant differences in COMT allelic and genotypic distribions were observed between patients with schizophrenia and healthy volunteers (x2 =8.16,11.74,Ps ＜ 0.05).The significant main effect of COMT genotype on habituation％ was observed (P ＜0.05) but no interaction genotype by diagnosis on the amplitude of auditory startle reflex,habituation％ and ％ PPI120 was observed (Ps ＞ 0.05).Conclusions:There may be a correlation between COMT genotype and adaptability,but not between COMT genotype and PPI deficit present in patients with schizophrenia

Epigenetics-based anticancer drugs:research advances / 国际药学研究杂志

In recent years,with the completion of the Human Genome Project and the development of mapping the Human Ge?nome Methylation Variable Site Map Plan,research on epigenetics and the generation and deveopment of cancer,epigenetic treatment drugs,especially the successful clinical application of the DNA methyltransferase and histone deacetylation inhibitors in the treatment of cancer patients,epigenetic has becoming a hot spot. This article reviews the recent progress in pharmacological action of epigenetics-based anticancer drugs,it may provide some new ideas to the therapy and fundamental research of cancer.

Epigenetics–based anticancer drugs: Research advances / 国际药学研究杂志

In recent years, with the completion of the Human Genome Project and the development of mapping the Human Genome Methylation Variable Site Map Plan, research on epigenetics and the generation and deveopment of cancer, epigenetic treatment drugs, especially the successful clinical application of the DNA methyltransferase and histone deacetylation inhibitors in the treatment of cancer patients,epigenetic has becoming a hot spot. This article reviews the recent progress in pharmacological action of epigenetics-based anticancer drugs, it may provide some new ideas to the therapy and fundamental research of cancer.

Effect of Polymorphisms of Three Genes Mediating Monoamine Signalling on Brain Morphometry in Schizophrenia and Healthy Subjects

OBJECTIVE: We examined the effect of risk alleles of polymorphisms of three schizophrenia risk genes that mediate monoamine signalling in the brain on regional brain volumes of schizophrenia and healthy control subjects. The risk alleles and the gene polymorphisms studied were: Val allele of catechol o-methyltransferase (COMT) rs4680 polymorphism; short allele of 5-hydroxy tryptamine transporter linked polymorphic region (5HTTLPR) polymorphism; and T allele of 5-hydroxy tryptamine 2A (5HT2A) rs6314 polymorphism. METHODS: The study was carried out on patients with recent onset schizophrenia (n=41) recruited from the outpatient department of National Institute of Mental Health and Neurosciences, Bangalore, India and healthy control subjects (n=39), belonging to South Indian Dravidian ethnicity. Individual and additive effects of risk alleles of the above gene polymorphisms on brain morphometry were explored using voxel-based morphometry. RESULTS: Irrespective of phenotypes, individuals with the risk allele T of the rs6314 polymorphism of 5HT2A gene showed greater (at cluster-extent equivalent to family wise error-correction [FWEc] p<0.05) regional brain volumes in the left inferior temporal and left inferior occipital gyri. Those with the risk alleles of the other two polymorphisms showed a trend (at p<0.001, uncorrected) towards lower regional brain volumes. A trend (at p<0.001, uncorrected) towards additive effects of the above 3 risk alleles (subjects with 2 or 3 risk alleles vs. those with 1 or no risk alleles) on brain morphology was also noted. CONCLUSION: The findings of the present study have implications in understanding the role of individual and additive effects of genetic variants in mediating regional brain morphometry in health and disease.

Correlation study on susceptible genes of obsessive compulsive disorder of Han population in northeast China / 中华行为医学与脑科学杂志

Objective To explore the relationship between Monoamine oxidase A (MAOA) receptor gene,catechol-O-methyltransferase (COMT) receptor gene,dopamine D3 receptor(DRD3) gene and 5-HT2C receptor gene(5-HT2c) of Han population in northern China and obsessive compulsive disorder.Methods Polymerase chain reaction amplification determination of MAOA-T1460C,COMT-Val158Met,DRD3-Ser9Gly,5-HT2c-C ys23Ser four loci receptor gene polymorphism in 164 patients with OCD patients including 103 core pedigrees of fragment length polymorphism,and association and linkage disequilibrium (TDT)analysis.Results There was no significant difference of MAOA-T1460C,COMT-Val158Met,DRD3-Ser9Gly,5-HT2c-Cys23Ser four receptor gene in the patient group and the control group of genotype and allele distribution difference(P＞0.05),four receptor gene loci were in accordance with the balance of the H-W,the MAOA-T1460C receptor gene in female patients group and control group,the early group and control group,which has forced thinking and difference of compulsive behavior group and the control group,only the obsessional group and the control group of genotype and allele distribution was statistically significant(P＜0.05),and the family group between chain(P=0.0001) ;5-HT2c-Cys23Ser receptor gene in the case group and the control group,male both forced thinking and compulsive behavior group and control group differences in genotype and allele distribution was statistically significant (P＜ 0.05),and between family groups exist chain (P=0.0389) ; COMT-Val158Met receptor gene in the control experiments were no significant difference(P＞0.05),and with the house group does not exist between the chain (P=0.0622) ;DRD3-Ser9Gly receptor gene in the control experiments were no significant difference(P＞0.05),and with the family groups there is no chain(P=0.1101).Conclusion MAOA-T1460C receptor gene polymorphism and 5-HT2c-Cys23Ser receptor gene polymorphisms may be the susceptible gene of obsessive compulsive disorder.

Determination of 4-Hydroxy-3-Methoxy Benzoic Acid by RP-HPLC and Inhibitory Effectof Phloroglucinol on Catecholamine-O-Methyl Transferase / 医药导报

Objective To detect content of 4-hydroxy-3-methoxy benzoic acid by RP-HPLC and observe the inhibitory effect of phloroglucinol on catecholamine-O-methyl transferase (COMT). Methods This study used the principle of 3,4-dihydroxy benzoic acid transforming to 4-hydroxy-3-methoxy benzoic acid under COMT’ s catalytic action. COMT (20 μL) was extracted from mouse liver homogenate. In a reaction system,10 μL catecol (1í10-3 mol·L-1 ) and 10 μL phloroglucinol (5í10-3 , 1í10-3 and 2í10-4 mol·L-1 ,respectively) were added. Products were determined by RP-HPLC to analyze effects of 4-hydroxy-3-methoxy benzoic acid on COMT. Results Phloroglucinol had inhibitory effect on COMT activity at concentrations of 5í10-3 mol·L-1 ,1í10-3 mol·L-1 and 2 í10-4 mol ·L-1 ,with the inhibition rate being 25. 3% ,17. 6% and 8. 9% ,respectively. Conclusion Phloroglucinol has an inhibitory effect on COMT activity,which is weaker than the effect of catechol of the same concentration.

Association of genetic variants of xenobiotic metabolic pathway with systemic lupus erythematosus.

In view of documented evidence that catechol estrogen-DNA adducts serve as epitopes for binding of anti-nuclear antibodies, genetic polymorphisms of the xenobiotic metabolic pathway involved in estrogen metabolism might contribute towards pathophysiology of systemic lupus erythematosus (SLE). To test this hypothesis, a case-control study was conducted. Cytochrome P 450 1A1 (CYP1A1) m4 (OR: 4.93, 95% CI: 1.31-18.49), catecholamine-o-methyl transferase (COMT) H108L (OR: 1.39, 95% CI: 1.03-1.88) and glutathione-S-transferase (GST) T1 null (OR: 1.83, 95% CI: 1.11- 3.01) variants showed association with SLE risk. SHEsis web-based platform analysis showed mild to moderate linkage disequilibrium between the CYP1A1 m1, m2 and m4 variants (D’: 0.19-0.37). Among the different haplotypes of CYP1A1, CAC-haplotype harboring CYP1A1 m1 variant showed association with SLE risk (OR: 1.46, 95% CI: 1.11-1.92). Multifactor dimensionality reduction analysis (MDR) showed potential gene-gene interactions between the phase II variants i.e. COMT H108L × GSTT1 null × GSTM1 null (p<0.0001) and also between the phase II and I variants i.e. COMT H108L × GSTT1 null × CYP1A1 m1 × CYP1A1 m2 in inflating the risk of SLE by 3.33-folds (95% CI: 2.30-4.82) and 4.00-folds (95% CI: 2.77-5.78), respectively. To conclude, hyperinducibility of CYP1A1 due to m1 and m4 variants and defective phase-II detoxification due to COMT H108L and GSTT1 null variants increase the susceptibility to SLE.

Impact of COMT H108L, MAOB int 13 A>G and DRD2 haplotype on the susceptibility to Parkinson’s Disease in South Indian subjects.

In view of documented evidence demonstrating the association of dopaminergic metabolism and neurotransmission with Parkinson’s disease (PD), a case-control study was conducted to investigate the impact of particular polymorphisms in the catechol O-methyl transferase (COMT) H108L, monoamine oxidase B (MAOB) int 13 A>G, dopamine transporter 1 (DAT1) A1215G, dopamine receptor D2 (DRD2) Taq1A, DRD2 Taq1B and DRD2 Taq1D genes on the susceptibility to PD. PCR-RFLP method was used for the genetic analysis. The COMT H108L polymorphism increased PD risk by 1.4-fold (95%CI: 1.02-1.98), whereas reduced risk was observed with MAOB int 13 A>G polymorphism (OR: 0.77, 95%CI: 0.51-0.99). Multifactor dimensionality reduction analysis showed gene-gene interactions between these two loci that resulted in loss of the protective role of MAOB G-allele in the presence of COMT L-allele. DAT1A1215G polymorphism in the exon 9 was not associated with PD. Individually, DRD2 polymorphisms showed null association. However, all-variant haplotype of DRD2 locus i.e. T-G-T haplotype showed 29.8-fold risk for PD compared to all-wild haplotype i.e., C-A-C haplotype (95%CI: 6.85-130.4). To conclude, genetic variants of COMT, MAOB and DRD2 loci modulate susceptibility to PD in South Indian subjects.

An association study on Gilles de la Tourette's syndrome and catechol-O-methyl transferase gene polymorphism / 中华行为医学与脑科学杂志

ObjectiveTo investigate the association between catechol-O-methyl transferase (COMT)gene polymorphism and Gilles de la Tourette' s syndrome(GTS).MethodsUsing Amplification Refractory Mutation System(ARMS) PCR genotyping assay method,a polymorphism (val158met) of COMT gene was genotyped in 112 of all GTS patients ( total GTS group) including 54 GTS-alone patients group,48 GTS + ADHD patients group among of them and 71 healthy controls.The correlation between positive association of polymorphism (val158met)of COMT gene in GTS and the age of onset in patients with GTS was also analyzed.ResultsCompared with healthy controls group,genotype of val158met did not differ in total GTS patients group or alone-GTS patients group (χ2 =0.56,P=0.756;χ2 =1.05,P=0.600 respectively).There was also no significant difference (P＞0.05)in allele distribution of val158met in total GTS patients group or alone-GTS patients group compared with controls group respectively (χ2 =0.18,P=0.669;χ2 =0.29,P=0.593 respectively).However,genotype distribution of val158met was significantly different between GTS + ADHD patients group and controls group( χ2 =6.35,P =0.041 ).The frequency of the val allele of this locus was significantly higher in GTS + ADHD patients group than those in controls group ( χ2 =5.49,P =0.019 ).The mean age of onset (6.80 ± 1.54 ) in 36 children within GTS + ADHD patients group with the val/val geantype of COMT gene val158met polymorphism was significantly earlier than the mean age of onset (8.04 ± 1.54)in 12 children in val/val genotype (P =0.016 ).ConclusionPolymorphism (val158met) of COMT gene may be associated with GTS children with comorbid ADHD,which may play an important role to make the age of onset in children with GTS become earlier.

Effects of trichostain A on multiple myeloma cell line U266 / 白血病·淋巴瘤

Objective To evaluate effects of trichostain A (TSA) on cell proliferation, cell cycles, apoptosis and invasiveness of multiple myeloma cell line U266; as well as active changes of methylation regulating proteins including DNA methyl-transferase(DNMTs), methyl-binding domain (MBD) proteins: MBD2 and MeCP2 after treated with TSA. Methods U266 cells were treated with different concentrations of TSA for 12, 24, 48 and 60 h. The proliferation activity of U266 cells was detected by MTT and the IC50 of 24 h was calculated. After U266 cells were treated with IC50, cell cycles were check out by dying with PI. mRNA of matrix metalloproteinase-2(MMP-2), bc1-2, bcl-xl and methylation regulating proteins (DNMTs, MBD2 and MeCP2) were detected by real-time PCR. FCM and Western blotting were used to measure expressions of MMP-2 and MBD2. Results MTT results revealed TSA inhibited proliferation of U266 cells in a dose-and time-dependent manner and the IC50 of 24 h was 0.07 μmol/L FCM analysis showed that TSA could arrest the cell cycle in G0/G1 and the proliferation index (PI) in U266 cells [(49.90 0.39)%]were significantly different after exposed to TSA (0.7 μmnol/L for 24h compared with that in the control cells[(55.78 0.49)%](P <0.01). After treated by TSA, the 2-△△Ct of MMP-2, bcl-2 and bcl-xl were 0.71 0.06, 5.04 0.92 and 2.95 0.35, respectively. There were great changes on mRNA of DNMT, MBD2 and MeCP2. TSA could reverse the transcription of DNMT, MBD2 and MeCP2. Conclusion TSA can arrest the U266 cell cycle in GVG, to prevent its proliferation and promote apoptosis, which maybe greatly connect with the changes of the methylation regulating proteins.

No Association between Val108/158Met Polymorphism on Catechol-O-Methyl Transferase(COMT) Gene and Smooth Pursuit Eye Movement(SPEM) Abnormality in Korean Schizophrenia Patients

OBJECTIVES: We investigated the association of Val108/158Met polymorphism on catechol-O-methyl transferase(COMT) gene with smooth pursuit eye movement(SPEM) abnormality in Korean schizophrenia patients. METHODS: We measured SPEM in 217 Korean schizophrenia patients(male 116, female 101) and divided them into two groups, one was a good SPEM function group and the other was a poor SPEM function group. Then we analyzed Val108/158Met polymorphism on COMT gene. We compared the differences of genotype and allele distributions of the polymorphism on COMT gene between the two groups. RESULTS: The natural logarithm value of signal/noise ratio(Ln S/N ratio) of the good SPEM function group was 4.39+/-0.33(mean+/-s.d.) and that of poor SPEM function group was 3.17+/-0.71. There were no statistically significant differences of age and male/female ratio between the two groups. There were no significant differences of genotype or allele distributions of the Val108/158Met polymorphism on COMT gene between the two schizophrenic groups. CONCLUSIONS: The results suggest that Val108/158Met polymorphism on COMT gene is not related to SPEM function abnormality in schizophrenia.

Molecular characterization of a cDNA encoding caffeoyl-coenzyme A 3 -O- methyltransferase of Stellaria longipes.

A cDNA clone encoding S-adenosyl-L-methionine: trans-caffeoyl-CoA 3-O-methyltransferase (EC 2·1·1·104; CCoAOMT) from Stellana longipes Goldie (long-stalked chickweed) was isolated and studied. Structural analysis of both the nucleotide sequence and the predicted amino acid sequence suggests that our cloned sequence encoded a CCoAOMT enzyme of Stellaria longipes, which shared overall structural similarity with other plant CCoAOMTs but exhibited certain distinct characteristics. Southern blot hybridization and cloning analyses indicating a small CCoAOMT gene family in the Stellana longipes genome and the absence of introns in the coding region of the cDNA- corresponding gene. Sequence variations in the coding region were found among three genotypes from geographically isolated populations. Higher levels of CCoAOMT mRNA were detected in stems and leaves than in roots. The cDNAencoded protein expressed in Eschendia coli was shown to utilize caffeoyl-CoA, but not caffeic acid or 5-hydroxy ferulic acid, as its substrate.

The effects of cromakalim on the mediator releases from guinea pig lung mast cell activated by specific antigen-antibody reactions

The inhibitory effect of cromakalim on the mediator release from mast cells caused by antigenantibody reactions was in controversy with the specific antigen used. However, it has recently been observed that cromakalim inhibits the release of mediators from superfused tracheal and parenchymal strips or lung mast cells after passive sensitization with the IgG1 antibody. An attempt, therefore, was made to determine the inhibitory mechanisms of cromakalim on the release of mediators such as histamine and leukotriene released by the activation of enzymes during mast cell activation. Guinea pig lung mast cells were purified through enzyme digestion, rough percoll and continuous percoll density gradients. The purified mast cells were prelabeled with [3H]palmitic acid. PLD activity was assessed more directly by the production of labeled phosphatidylethanol by PLD-mediated transphosphatidylation in the presence of ethanol. In the cells labelled with [3H]myristic acid, [3H] DAG production was measured. The methyltransferase activity was assessed by measuring the incorporation of [3H]methyl moiety into phospholipids in sensitized mast cells labelled with L-[3H] methylmethionine. cAMP level was measured by radioimmunoassay. Cromakalim resulted in a decrease in the amount of histamine and leukotrienes releases by 30% in the ovalumin-induced mast cell. Cromakalim had little effect on phospholipase D activity enhanced by the activated mast cell. Cromakalim inhibited the initial increase of diacylglycerol production during mast cell activations. Cromakalim inhibited the phospholipid methylation increased in the activated mast cell. These results show that cromakalim decreases histamine release by inhibiting the initial increase of 1,2-diacylglycerol during the mast cell activation, which is mediated via the phosphatidylinositide-phospholipase C system rather than the phosphatidylcholine-phospholipase D system. Furthermore, cromakalim reduces phosphatidylcholine production by inhibiting the methyltransferase, which decreases the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrienes.

The effects of cromakalim on the mediator releases from guinea pig lung mast cell activated by specific antigen-antibody reactions

The inhibitory effect of cromakalim on the mediator release from mast cells caused by antigenantibody reactions was in controversy with the specific antigen used. However, it has recently been observed that cromakalim inhibits the release of mediators from superfused tracheal and parenchymal strips or lung mast cells after passive sensitization with the IgG1 antibody. An attempt, therefore, was made to determine the inhibitory mechanisms of cromakalim on the release of mediators such as histamine and leukotriene released by the activation of enzymes during mast cell activation. Guinea pig lung mast cells were purified through enzyme digestion, rough percoll and continuous percoll density gradients. The purified mast cells were prelabeled with [3H]palmitic acid. PLD activity was assessed more directly by the production of labeled phosphatidylethanol by PLD-mediated transphosphatidylation in the presence of ethanol. In the cells labelled with [3H]myristic acid, [3H] DAG production was measured. The methyltransferase activity was assessed by measuring the incorporation of [3H]methyl moiety into phospholipids in sensitized mast cells labelled with L-[3H] methylmethionine. cAMP level was measured by radioimmunoassay. Cromakalim resulted in a decrease in the amount of histamine and leukotrienes releases by 30% in the ovalumin-induced mast cell. Cromakalim had little effect on phospholipase D activity enhanced by the activated mast cell. Cromakalim inhibited the initial increase of diacylglycerol production during mast cell activations. Cromakalim inhibited the phospholipid methylation increased in the activated mast cell. These results show that cromakalim decreases histamine release by inhibiting the initial increase of 1,2-diacylglycerol during the mast cell activation, which is mediated via the phosphatidylinositide-phospholipase C system rather than the phosphatidylcholine-phospholipase D system. Furthermore, cromakalim reduces phosphatidylcholine production by inhibiting the methyltransferase, which decreases the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrienes.

Expression of DNMT1,?-catenin and P-GSK-3? in colon carcinoma / 西安交通大学学报(医学版)

Objective To examine DNMT1,?-catenin and P-GSK-3? expressions in tissues of colon carcinoma compared with tumor clinicopathological parameters including differentiation and metastasis.Methods SYBR Green real-time PCR and immunoblotting method were used to detect these gene expressions in 62 tissues of colon carcinoma and 21 tissues of normal colon.Results Overexpressions of DNMT1,?-catenin,and P-GSK-3? were found in colon carcinoma tissues.The expression of DNMT1 was found to be higher in poorly differentiated tissues.The results also demonstrated statistical significance in the expression of ?-catenin involving differentiation and metastasis,but no statistical significance in the expression of P-GSK-3?.Conclusion These results show that DNMT1,?-catenin and P-GSK-3? expressions are regulated throughout colon cancer progression.