The purpose of this study is to obtain the clinicopathological characteristics of replication error-positive (RER ) gastric adenocarcinoma in Korean, and to identify the significance of RER in adenoma stage of gastric carcinogenesis. Microsatellite instability was examined at D2S71, D2S119, D3S1067, D6S87, D11S905, DM, AR, VWF, HPRT, and BAT-26 loci. Frameshift mutation of BAX gene was analyzed in RER tumors. Normal and tumor DNA of 76 cases of gastric carcinoma and 25 cases of adenoma were examined. RER was found in 8 of 76 cases (10.5%), and it was more frequently found in adenocarcinoma of female (17.7%) than those of male (4.8%). The frequency of RER was not different between the histologic types, age of the patient, anatomical location of the carcinoma, and the stage. The RER found in adenoma suggests that RER contributes to the malignant transformation early in the adenoma stage of the gastric carcinogenesis. None of the RER tumors revealed frameshift mutation of the BAX gene.
Adenocarcinoma* ; Adenoma ; Carcinogenesis ; DNA ; Female ; Frameshift Mutation ; Humans ; Hypoxanthine Phosphoribosyltransferase ; Male ; Microsatellite Instability

OBJECTIVE: It has been known that the enzyme deficiency associated with uric acid production is the minor cause of gout. The purpose of this study is to evaluate the role of hypoxanthine- guanine phosphoribosyltransferase (HGPRT) activity deficiency in the development of gout. METHODS: Assay of HGPRT activity was performed on 38 gout patients and 107 controls compromising 82 men and 25 women. In the patients with gout, age of onset, sex, family history of gout, duration of disease, serum uric acid level, and uric acid concentration of 24-hour urine were analyzed. RESULTS: In gout patients and control male group, the HGPRT activity was lowest in the age of forties. But, in female, there was no decreasing tendency with aging. In 4 gout patients, HGPRT activities were severely deficient and their ages of onset were less than 30 years old, and all have a family history of gout. CONCLUSION: In men, the lowest HGPRT activity in the age of forties could be one of the factors that make the onset of gout is highest in the fifth decade of age. The deficiency of HGPRT enzyme and family history of gout are the important factors in the pathogenesis of early onset gout.
Adult ; Age of Onset ; Aging ; Female ; Gout* ; Humans ; Hypoxanthine Phosphoribosyltransferase* ; Male ; Uric Acid

Deficiency of hypoxanthine-guanine phosphoribosyltransferase is a purine nucleotide disorder and is the most common genetic cause of uric acid overproduction. This disease has a wide range of spectrum with regard to neurological features depending on the extent of the enzymatic deficiency. Complete deficiency of hypoxanthine-guanine phosphoribosyltransferase, called Lesch-Nyhan syndrome, is presented with hyperuricemia and characteristic neurological manifestation and self-mutilation. Partial hypoxanthine-guanine phosphoribosyltransferase--deficient patients are presented with a various intensities of the aforementioned symptoms, from almost normal neurologic manifestation to a severe form along with hyperuricemia. We report a twenty-year-old man with complete hypoxanthine-guanine phosphoribosyltransferase mutation and Lesch-Nyhan sydrome, who manifested gouty arthritis without neurologic symptom.
Arthritis, Gouty* ; Humans ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Lesch-Nyhan Syndrome* ; Neurologic Manifestations* ; Uric Acid

Animals ; Genes, T-Cell Receptor ; genetics ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Mice ; Mutation

OBJECTIVE: To carry out genetic testing and prenatal diagnosis for a Chinese pedigree with Lesch-Nyhan syndrome (LNS) but no specimen from the affected probands. METHODS: All affected individuals in this pedigrees were male and had deceased during childhood, with no biological specimen left. Based on their typical neurological dysfunction and tendency for self-mutilation, the diagnosis of LNS was suspected. Sanger sequencing was carried out to detect potential variant of the HPRT1 gene among female members from the pedigree. Following the identification of the pathogenic variant, prenatal diagnosis was provided for a high-risk fetus. RESULTS: The proband's mother and three other females were found to harbor heterozygous c.500_501delGGinsC (p.Arg167fs*23) variant of the HPRT1 gene, which was unreported previously. Prenatal diagnosis showed that the fetus was a male and had inherited the same pathogenic variant. CONCLUSION The c.500_501delGGinsC variant of the HPRT1 gene probably underlay the LNS in this pedigree. Above finding has provided a basis for prenatal diagnosis and genetic counseling for this pedigree.
Male ; Female ; Humans ; Pregnancy ; Lesch-Nyhan Syndrome/genetics* ; Pedigree ; Hypoxanthine Phosphoribosyltransferase/genetics* ; Prenatal Diagnosis ; China ; Mutation

Lesch-Nyhan disease is a very rare X-linked recessive disorder characterized by mental retardation, spasticity resembling cerebral palsy, choreoathetosis, self-mutilation and hyperuricemia. Self-mutilative behavior is a hallmark of the disease. The underlying defect is a deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT). We report on a fourteen-year-old boy, who manifested gouty arthritis and mild renal insufficiency with Lesch-Nyhan disease, lacking self-mutilative behavior in spite of undetectable HPRT activity. Though there were several reports about some cases of Lesch-Nyhan disease in the past Korean literature, the cases were classic forms with definite neurological manifestation. As far as we know, this is the first case of Lesch-Nyhan disease without self-mutilation in Korea.
Arthritis, Gouty ; Cerebral Palsy ; Gout ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Intellectual Disability ; Korea ; Lesch-Nyhan Syndrome ; Muscle Spasticity ; Neurologic Manifestations ; Renal Insufficiency

BACKGROUND: Quantitative real-time polymerase chain reaction (qPCR) is the most reliable tool for gene expression studies. Selection of housekeeping genes (HKGs) that are having most stable expression is critical to carry out accurate gene expression profiling. There is no 'universal' HKG having stable expression in all kinds of tissues under all experimental conditions. METHODS: The present study aims to identify most appropriate HKGs for gene expression analysis in glioblastoma (GBM) samples. Based on literature survey, six most commonly used HKGs that are invariant in GBM were chosen. We performed qPCR using RNA from formalin fixed paraffin embedded GBM samples and normal brain samples to investigate the expression pattern of HPRT, GAPDH, TBP, B2M, B2M, RPL13A, and RN18S1 with different abundance. A simple Deltacycle threshold approach was employed to calculate the fold change. RESULTS: Our study shows that the expression of RPL13A and TBP were found to be most stable across all the samples and are thus suitable for gene expression analysis in human GBM. Except for TBP, none of the other conventionally used HKGs in GBM studies e.g., HPRT and GAPDH were found to be suitable as they showed variation in RNA expression. CONCLUSION: Validation of HKGs is therefore immensely specific for a particular experimental setup and is crucial in assessing any new setup.
Brain ; Formaldehyde ; Gene Expression Profiling ; Gene Expression* ; Genes, Essential* ; Glioblastoma* ; Humans ; Hypoxanthine Phosphoribosyltransferase ; Paraffin ; Real-Time Polymerase Chain Reaction* ; RNA

OBJECTIVETo study genetic damage of workers alone occupationally exposed to methotrexate (MTX) with three end-points.

METHODSThe blood samples from 21 workers exposed to MTX and 21 controls were detected with micronucleus test, comet assay, hprt gene mutation test and TCR gene mutation test.

RESULTSThe mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in 21 workers were 10.10 per thousand +/- 0.95 per thousand and 8.05 per thousand +/- 0.75 per thousand, respectively, which were significantly higher than those (5.48 per thousand +/- 0.82 per thousand and 4.38 per thousand +/- 0.58 per thousand) in control (P < 0.01). The mean tail length (MTL) of 21 workers and 21 controls were (1.30 +/- 0.06) microm and (0.07 +/- 0.01) microm, respectively, there was significant difference between workers and controls (P < 0.01). But the difference between workers and controls for mean tail moment (MTM) was not significant (P > 0.05). The average mutation frequency (Mf-hprt) of hprt and (Mf-TCR) of TCR in workers were 1.00 per thousand +/- 0.02 per thousand and (6.87 +/- 0.52) x 10(-4), respectively, which were significantly higher than those [0.86 per thousand +/- 0.01 per thousand and (1.67 +/- 0.14) x 10(-4)] in control (P < 0.01).

CONCLUSIONThe genetic damage to some extent appeared in workers occupationally exposed to methotrexate.

Adult ; Comet Assay ; DNA Damage ; Female ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Male ; Methotrexate ; toxicity ; Micronucleus Tests ; Middle Aged ; Mutation ; Occupational Exposure

Programmed cell death (PCD), or apoptosis, plays an important role in embryonic development, metamorphsis, hormone-dependent atrophy and tumor growth, as a physiological event regulating the cell number or eliminating damaged cells. Currently hybridoma cell production, resulting from the fusion of myeloma cells with antibody producing spleen cells, is widely used in various fields of life science. This technique requires a hypoxanthine guanine phosphoribosyl transferase (HGPRT) deficient mutant myeloma cell line as a fusion partner. When these rnutant cells are treated with aminopterin plus hypoxanthine-thymidine (HAT) after the cell fusion they are selectively and efficiently eliminated so that one could get only fused hybridoma cells. But there hasn't been any report regarding this selective elimination mechanism of HGPRT mutant myeloma cell. By using HGPRT myeloma V653 as a model system this study demonstrated that PCD was induced by aminopterin treatment of this V653 cell. And this PCD was further characterized by cotreatement of cycloheximide, actinomycin-D, and calcium ionophore A23187 together with aminopterin. The apoptotic endonuclease involved in this PCD process was also detected and characterized. When V653 cells were incubated for the various periods of time in the presence of 0.4 uM aminopterin, the viability was continued to decrease until 48 hours of aminopterin treatment and there was no viable cell affer 36 hours of incubation. DNA fragmentation was detectable 3 hours of incubation and peaked between 12 and 18 hours of aminopterin treatment. The induction of cell death and DNA fragmentation of V653 cells by aminopterin were inhibited by protein synthesis inhibitor, cycloheximide, and RNA synthesis inhibitor, actinomycin-D and maximal inhibitory effects on cell death were seen at concentrations of 2 ug/ml and 0.5 uM, respectively. Ca2+ ionophore A23187 promoted aminopterin-induced cell death of V653. When the cells were coincubated with A23187 in the presence of aminopterin, cell viability was remarkably decreased at concentrations of more than 2 uM and DNA fragmentation was increased at concentrations of more than 0.2 uM. A23187 also induced cell death when the cells were treated with A23187 alone. When endogenous endonuclease activities of nuclei isolated from intact healthy cells and aminopterin-treated cells were compared for four different conditions, there were notable increases in the Ca2+/Mg2+ -independent and the Mg2+ -dependent endonuclease activity after incubation with aminopterin for 12 hours. In northern blot analysis, induction of c-myc gene was observed in aminopterin-treated V653 cell reached peak at 2 hours and thern decreased drastically.
Aminopterin* ; Animals ; Apoptosis* ; Atrophy ; Biological Science Disciplines ; Blotting, Northern ; Calcimycin ; Calcium ; Cell Count ; Cell Death ; Cell Fusion ; Cell Line ; Cell Survival ; Cycloheximide ; DNA Fragmentation ; Embryonic Development ; Female ; Genes, myc ; Guanine ; Hybridomas ; Hypoxanthine ; Hypoxanthine Phosphoribosyltransferase ; Mice* ; Pregnancy ; RNA ; Spleen ; Transferases

Lesch-Nyhan syndrome is an X-linked recessive disorder characterized by hyperuricemia, choreoathetosis, spasticity, mental retardation, and compulsive, self-injurious behavior. This disorder results from a complete deficiency of the purine salvage enzyme, hypoxanthine-guanine phosphoribosyl transferase(HPRT). We report here on a case of Lesch-Nyhan syndrome in a 1-year, 7-month-old male who presented with frequent vomiting, failure to thrive, and developmental delay. The diagnostic work-up revealed hyperuricemia, hyperuricosuria, and medullary nephrolithiasis. The HPRT activity in the erythrocytes was undetectable with a biochemical assay. We also identified de novo mutation which was a deletion of the 649th base, adenosine, in HPRT gene(649delA) by analysis of cDNA using RT-PCR technique coupled with direct sequencing.
Adenosine ; DNA, Complementary ; Erythrocytes ; Failure to Thrive ; Humans ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Infant ; Intellectual Disability ; Lesch-Nyhan Syndrome* ; Male ; Muscle Spasticity ; Nephrolithiasis ; Self-Injurious Behavior ; Vomiting