No Abstract Available.
Histidine*

No Abstract Available.
Histidine*

To investigate the involvement of expression of the Fragile Histidine Triad(FH1T) gene product in the process of carcinogenesis and progression in cervical carcinoma, we examined its expression by immunohistochemical method in 15 cervical invasive carcinomas, 10 low grade cervical intraepithelial neoplasias(CINs) and 30 high grade CINs(CMI and III). We detected expression of FHIT gene product in 4 of 15(27%) of invasive carcinomas, 3 of 10(30%) low grade CIN and 7 of 30(23%) of high grade CIN, while we detected expression of FHIT gene product in 28 of 45(62%) normal and metaplastic epithelium near the tumor. Thesc data indicate that loss of expression of FH1T gene product has some role in the early tumorigenesis of uterine cervical carcinoma, but not the consequence of the pregression of the tumor.
Carcinogenesis ; Epithelium ; Histidine* ; Immunohistochemistry

Ergothioneine (ERG) is a natural antioxidant that has been widely used in the fields of food, medicine and cosmetics. Compared with traditional plant extraction and chemical synthesis approaches, microbial synthesis of ergothioneine has many advantages, such as the short production cycle and low cost, and thus has attracted intensive attention. In order to engineer an ergothioneine high-yielding Escherichia coli strain, the ergothioneine synthesis gene cluster egtABCDE from Mycobacterium smegmatis and egt1 from Schizosaccharomyces pombe were introduced into E. coli BL21(DE3) to generate a strain E1-A1 harboring the ergothioneine biosynthesis pathway. As a result, (95.58±3.2) mg/L ergothioneine was produced in flask cultures. To further increase ergothioneine yield, the relevant enzymes for biosynthesis of histidine, methionine, and cysteine, the three precursor amino acids of ergothioneine, were overexpressed. Individual overexpression of serA^T410STOP and thrA resulted in an ergothioneine titer of (134.83±4.22) mg/L and (130.26±3.34) mg/L, respectively, while co-overexpression of serA^T410STOP and thrA increased the production of ergothioneine to (144.97±5.40) mg/L. Eventually, by adopting a fed-batch fermentation strategy in 3 L fermenter, the optimized strain E1-A1-thrA-serA^* produced 548.75 mg/L and 710.53 mg/L ergothioneine in glucose inorganic salt medium and rich medium, respectively.
Culture Media ; Ergothioneine/metabolism* ; Escherichia coli/metabolism* ; Fermentation ; Histidine/metabolism* ; Metabolic Engineering

BACKGROUND: Recent genome-wide sequencing studies have identified unexpected genetic alterations in cancer. In particular, missense mutations in isocitrate dehydrogenase-1 (IDH1) at arginine 132, mostly substituted into histidine (IDH1-R132H) were observed to frequently occur in glioma patients. METHODS: We have purified recombinant IDH1 and IDH1-R132H proteins and monitored their catalytic activities. In parallel experiments, we have attempted to find new selective IDH1-R132H chemical inhibitor(s) from a fragment-based chemical library. RESULTS: We have found that IDH1, but not IDH1-R132H, can catalyze the conversion of isocitrate into alpha-ketoglutarate (alpha-KG). In addition, we have observed that IDH1-R132H was more efficient than IDH1 in converting alpha-KG into (R)-2-hydroxyglutarate (R-2HG). Moreover, we have identified a new hit molecule, e.g., 2-(3-trifluoromethylphenyl)isothioazol-3(2H)-one as a new selective IDH1-R132H inhibitor. CONCLUSIONS: We have observed an underlying biochemical mechanism explaining how a heterozygous IDH1 mutation contributes to the generation of R-2HG and increases cellular histone H3 trimethylation levels. We have also identified a novel selective IDH1-R132H chemical hit molecule, e.g., 2-(3-trifluoromethylphenyl)isothioazol-3(2H)-one, which could be used for a future lead development against IDH1-R132H.
Arginine ; Glioma ; Histidine ; Histones ; Humans ; Mutation, Missense

OBJECTIVE: To evaluate Fragile histidine triad (Fhit) and p53 expression pattern in cervical intraepithelial neoplasm (CIN) and invasive cervical cancer, and to verify the correlation between the loss of Fhit and clinicopathological parameters of invasive cervical carcinoma and the relationship between Fhit and p53 expression. METHODS: 10 low-grade squamous intraepithelial lesions (LSIL), 16 high-grade squamous intraepithelial lesions (HSIL), and 21 invasive cervical carcinomas were evaluated by immunohistochemical staining for Fhit and p53 primary antibody. Their expression patterns in CIN and invasive cervical cancer were analysed semiquantitatively as positive and negative by the staining area and intensity. Clinicopathological data were obtained by review of patients' hospital records. RESULTS: Compared with CIN (LSIL and HSIL), invasive cervical carcinoma showed significantly loss of Fhit expression (p<0.05). P53 expression did not show the significant difference between CIN and invasive cervical cancer. There was no relationship between loss of Fhit and p53 expression in CIN and invasive cervical cancer. But loss of Fhit expression in invasive cervical cancer was also significantly associated with FIGO stage (p<0.05). CONCLUSION: Our results suggest that loss of Fhit expression may play an important role in the malignant transformation of CIN to invasive cancer. However, further molecular studies are needed to elucidate the role of Fhit gene in the carcinogenesis of cervical cancer.
Carcinogenesis ; Cervical Intraepithelial Neoplasia* ; Histidine ; Hospital Records ; Uterine Cervical Neoplasms*

PURPOSE: Genomic alterations and abnormal expression of the fragile histidine triad (FHIT) gene in gastric cancer were examined to determine whether the FHIT gene is actually a frequent target for alteration during gastric carcinogenesis. MATENRIALS AND METHODS: To correlate DNA and RNA lesions of the FHIT gene with the effect on FHIT protein expression, in 40 gastric cancers, we investigated the FHIT gene for loss of heterozygisity (LOH), aberrant transcripts, and protein expression. RESULTS: Allelic loss at D3S1300 was detected in 7 of 38 (19%) informative cases. Aberrant transcripts were observed in 20 of 40 (50%) cases. Significant reduction of FHIT protein expression was observed in 22 of 40 (55%) cases. Aberrant FHIT transcription was shown to be associated with loss of FHIT protein expression. However, aberrent FHIT transcripts themselves were not associated with any clinicopathological parameters, such as age, sex, tumor site, or clinical stage. Moreover, there was no association between the presence of LOH at D3S1300 and the expression of aberrant FHIT transcripts. CONCLUSION: The high frequency of aberrant FHIT transcripts, the significant rate of LOH at D3S1300, and the altered expression of the FHIT protein indicate that alterations of the FHIT gene can play an important role in gastric carcinogenesis.
Carcinogenesis ; DNA ; Gene Expression* ; Histidine* ; Loss of Heterozygosity ; RNA ; Stomach Neoplasms*

OBJECTIVE: To investigate the expression of fragile histidine triad (FHIT) protein and the possible relationship between FHIT expression and clinicopathological indices in cervical adenocarcinoma. METHODS: FHIT protein expression was examined in 40 cases of cervical adenocarcinoma stage Ia to IIa and 28 cases of corresponding normal endocervical tissue by immunohistochemical method. We analyzed the relationship between the reduction of FHIT protein expression and several prognostic factors such as histological grade, lymph node metastasis, tumor size, cervical invasion depth and parametrial invasion. We used Fisher's exact test for statistical analysis. RESULTS: The FHIT protein expression was positive in 77.5% (31/40) of cervical adenocarcinoma tissue, and reduced its expression in 22.5% (9/40) whereas positive in 100% (28/28) cases of adjacent normal endocervical gland. The FHIT expression was decreased in 14.3% (4/28) of cancers without lymph node metastasis but 55.5% (5/9) of those with metastasis (p=0.023). And the reduction of FHIT expression was found in 34.8% (8/23) of grade II and III cancers and only 6.3% (1/16) of grade I (p=0.056). CONCLUSION: Loss of FHIT protein expression may be associated with metastasis and poor prognosis of cervical adenocarcinoma.
Adenocarcinoma* ; Histidine ; Immunohistochemistry ; Lymph Nodes ; Neoplasm Metastasis ; Prognosis

The aim of this study was to express and purify human histydyl-tRNA synthetase related gene and to prepare its polyantibody. The open reading frame was amplified by PCR, and then recombined into prokaryotic expression vector pQE30 and transformed into E. coli M15 for expression. The expressed products were induced by IPTG after the reconstructed pQE30 was transferred into M15. After purified by Ni affinity chromatography, the product was identified to be a single band by SDS-PAGE. The rabbits were inoculated with purified products. High-titer polyantibody was successfully prepared. Highly-purified expression product and prepared polyantibody may provide a good basis for further study.
Antibodies/*genetics ; Antibodies/immunology ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Histidine-tRNA Ligase/biosynthesis ; Histidine-tRNA Ligase/*genetics ; Histidine-tRNA Ligase/immunology ; Open Reading Frames/genetics ; Prokaryotic Cells/metabolism

OBJECTIVE: The abnormal expression of fragile histidine triad (FHIT) gene has been frequently reported in a variety of epithelial malignancies including cervical carcinoma. Furthermore, in a recent study it was proposed that transcriptional inactivation of FHIT, as a consequence of aberrant 5'-CpG island methylation, plays an important role in the carcinogenesis of human cervical carcinoma. The authors sought to determine whether abnormal FHIT transcription occurs in human cervical carcinoma, and if so, whether this abnormal expression is associated with aberrant 5'-CpG island methylation. In addition, the clinical significance of FHIT inactivation was investigated in Korean women with cervical cancer. METHODS: To examine for abnormal transcripts of the FHIT gene, quantitative RT-PCR, genomic DNA-PCR and nonisotopic RT-PCR-SSCP analysis were performed using the standard method. The methylation status was determined by methylation specific PCR and bisulfite DNA sequencing. RESULTS: The FHIT gene was down-regulated in 15 of 58 (25.9%) cervical carcinomas. FHIT promoter hypermethylation was detected in 15 of 15 (100%) abnormally expression in cervical carcinomas. Bisulfite DNA sequencing confirmed these findings and a significant correlation was found between CpG site hypermethylation and low FHIT expression. However, no significant correlation was found between reduced FHIT expression and clinicopathological characteristics. CONCLUSION: In this study, FHIT inactivation in cervical cancer was found to be strongly correlated with 5'-CpG island hypermethylation rather than a genetic alteration. Furthermore, no significant relation was found between a lack of FHIT expression and the prognostic factors of cervical cancer in our Korean cohort.
Cohort Studies ; Female ; Histidine ; Humans ; Methylation ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sulfites ; Uterine Cervical Neoplasms