A case of neonatal diabetes mellitus is described. The child presented with low birth weight but was normal in appearance. She was acidotic and ketonuria was observed. The HLA typing was DR1 and 3, and insulin autoantibodies were negative. Genetic analysis with polymorphic DNA markers for chromosome 6 indicated biparental inheritance. She required insulin therapy for the control of hyperglycemia, and insulin dependence continues after 8 months of age.
Autoantibodies ; Child ; Chromosomes, Human, Pair 6 ; Diabetes Mellitus* ; Genetic Markers ; Histocompatibility Testing ; Humans ; Hyperglycemia ; Infant, Low Birth Weight ; Infant, Newborn ; Insulin ; Ketosis ; Wills

BACKGROUNDS: The molecular genetic characteristcs of cis-AB blood group have shown that its allele had C, G, C and C at nucleotide positions (nps) 526, 703, 796 and 803, respectively. And all cis-AB analysed and reported molecular genetically in Korea and Japan had T at np 467 (leucine at amino acid position 156). We report a first case of cis-AB with C at np 467 (proline at amino acid position 156). METHODS: Genomic DNA was extracted from peripheral blood of cis-AB patient and amplified by DS1/DS2 and DS3/DS4 allele-specific primers. After PCR, we analysed nps 261, 467, 526, 646, 703, 796, and 803 by restriction digestion, autoradiography and automatic sequencing. RESULTS: PCR-RFLP with DS1/DS2 primers and restriction enzyme KpnI showed that cis-AB had an 0 allele. The results of genomic sequencing, autoradiography and restriction digestion showed that cis-AB allele at nps 467, 526, 646, 703, 796 and 803 had C, C, T, G, C and C, respectively. CONCLUSION: This cis-AB showed characteristic molecular genetic features at nps 526, 703, 796, and 803. And this is a first case of A(Pro) cis-AB with C at np 467. (Korean J Blood Transfusion 10(1): 13-19, 1999)
Alleles ; Autoradiography ; Blood Transfusion ; Cytosine* ; Digestion ; DNA ; Humans ; Japan ; Korea ; Molecular Biology* ; Polymerase Chain Reaction

No abstract available.
Genetic Therapy*

PURPOSE: Ataxia Telangiectasia (AT) is a hereditary multi-systemic disease resulting from mutations of AT gene and is characterized by progressive neurodegeneration, cancer, immune system defects, and hypersensitivity to ionizing radiation. AT gene has a homologue sequence of PI3-kinase. The activity and cellular function of PI3-kinase in AT gene remains unclear. This study was undertaken to evaluate the function of AT gene through the effect on cell survival and differentiation by the inhibition of AT gene expression. MATERIALS AND METHODS: NH2-terminal portion of AT gene was isolated from MCF-7 cells by RT-PCR. The isolated DNA fragment was ligated in reverse orientation in pcDNA3. This antisense ATM expression vector was transfected to PC-12 cells by calcium phosphate method, and the transformed cells were selected using G418 and immunohisto- chemistry. To analyze the cell survival and differentiation, cells were cultured in serum free medium supplemented with/without NGF. We performed the immunoprecipitation for the p53 induction of cells after ionizing radiation, and the FACS for the apoptosis of cells after the exposure of wortmanin. RESULTS: PC-12 cells which down-regulated AT gene (like ATM, AT mutated) showed decreased survival and ceased differentiation with NGF. Also, PC-12 (ATM) cells showed increased apoptosis with wortmanin and reduced or delayed p53 induction after ionizingradiation. CONCLUSION: Results obtained from these studies suggest that AT gene regulates survival and differentiation of PC-12 cells through PI3-kinase activity. It seems that apoptosis is induced by the inhibition of AT gene expression.
Apoptosis ; Ataxia Telangiectasia ; Calcium ; Cell Survival ; Chemistry ; DNA ; Gene Expression ; Hypersensitivity ; Immune System ; Immunoprecipitation ; MCF-7 Cells ; Nerve Growth Factor ; Phenotype* ; Phosphatidylinositol 3-Kinases ; Radiation, Ionizing

The authors regret that two co-authors were missing in the article.

PURPOSE: Cefodizime is a new third-generation cephalosporin which has a structure and immunomodutation properties similar to cefotaxime. Various studies on cefodizime have demonstrated the direct eradication of bacteria in cooperation with the host defense mechanism, particularly with phagocytosis. We evaluated the effects of cefodizime on the phagocytosis of COS-1 cells transfected with FcgammaRI/gammagamma or FcgammaRIIA cDNA. METHODS: Phagocytosis was measured using the in vitro COS-1 cell modeling system according to Schreiber's method. COS-1 cells, which lack endogenoous Fcgammareceptors but have phagocytic potential, were transfected with either FcgammaRI/gammagammaor FcgammaRIIA cDNA. COS-1 cells, as target cells, were treated with antibiotics for 1 or 24 hours and incubated for 30 min with IgG coated sheep RBCs. Adhered IgG coated sheep RBCs were removed after brief exposure to hypotonic phosphate buffered saline. Phagocytosis index (PI) was calculated as the number of ingested RBCs per 100 phagocytic cells after wright-Giemsa staining. RESULTS: COS-1 cells tranfected with FcgammaR (either FcgammaRI/gammagamma or FcgammaRIIA cDNA) showed the phagocytic activity against IgG coated sheep RBC, while untransfected COS-1 cells did not. After treatment with cefodizime, phagocytic activity of FcgammaRI/gammagammacDNA transfected COS-1 cells was significantly increased, while that of FcgammaRIIA cDNA transfected COS-1 cells did not. Marked enhancement of phagocytosis of COS-1 cells was observed after treatment with cefodizime, but was not observed with ceftriaxone or moxalactam. CONCLUSION: Cefodizime showed marked enhancement of phagocytic activity of FcgammaR transfected COS-1 cells. FcgammaRI seems to play an important role in the enhancement of phagocytosis. Further studies will be required.
Animals ; Anti-Bacterial Agents ; Bacteria ; Cefotaxime ; Ceftriaxone ; COS Cells ; DNA, Complementary ; Immunoglobulin G ; Moxalactam ; Phagocytes ; Phagocytosis* ; Sheep

PURPOSE: Insulin-like growth factor binding protein 3 (IGFBP-3) binds to IGF and acid labile subunit in blood, that regulates the action of IGF by modulating the interaction of IGF with the IGF receptor. Recent studies have shown that IGFBP-3 levels are decreased in prostate cancer patients. We examined IGFBP-3 profiles in prostate cancer patients and determined the effect of treatment on serum IGFBP-3 level in those patients. MATERIALS AND METHODS: Control groups were composed of age-matched healthy subjects and patients with benign prostatic hyperplasia (BPH). Patients with prostate cancer were divided into localized, locally advanced, metastatic and hormone-refractory. Serum samples were collected and stored at 70oC until use. The IGFBP-3 profile was measured by Western ligand blots. RESULTS: The serum IGFBP-3 level in patients with prostate cancer was significantly lower than healthy subjects or patients with BPH. Following the treatment of prostate cancer, IGFBP-3 was increased compared to that seen in pretreated prostate cancer. In hormone-refractory prostate cancer, IGFBP-3 was decreased again. However, there was no correlation between IGFBP-3 and tumor stage or Gleason score. CONCLUSIONS: These data show that IGFBP-3 levels are decreased in pretreated and hormone-refractory prostate cancer. Our results demonstrate that serum IGFBP-3 may be a useful marker in the detection and management of prostate cancer.
Diagnosis* ; Humans ; Insulin-Like Growth Factor Binding Protein 3* ; Neoplasm Grading ; Prostate* ; Prostatic Hyperplasia ; Prostatic Neoplasms*

PURPOSE: Gap junction intercellular communication(GJIC) is an important mechanism of the bystander effect in herpes simplex thymidine kinase/ganciclovir(HSVtk/GCV) gene therapy Therefore, we attempted to enhance the bystander effect in vitro by exogenous overexpressing connexin 37(Cx37) in cells to increase GJIC. METHODS: NIH3T3 cells were transfected with the Cx37 and HSVtk gene or the HSVtk gene alone by the calcium phosphate method, and we detected their expression from these cells by RT-PCR. GCV-mediated cytotoxicity and the bystander effect of each transfectant was then assessed and compared. RESULTS: Cells transfected with HSVtk became sensitive to low concentration of GCV. We found significantly increased cytotoxicity in HSVtk/GCV gene therapy after introduction of the HSVtk and Cx37 genes together compared with the cytotoxicity seen after introduction of the HSVtk gene in vitro. Co-expression of the HSVtk and Cx37 genes potentiates HSVtk/GCV gene therapy through the bystander effect. CONCLUSION: These results indicated that the increase of GJIC using Cx37 have potentiated the by stander effect of HSVtk/GCV therapy, and may be a new approach to improve response in suicidal cancer gene therapy.
Bystander Effect* ; Calcium ; Gap Junctions* ; Genes, Neoplasm ; Genetic Therapy* ; Herpes Simplex ; Thymidine

PURPOSE: Fusion genes(EWS-Fli-1 and EW.S-erg) function as transcription activators and are essential for maintaining tumorigenic properties in Ewing's sarcoma cells. Several reports have noted that Ets family transcription factors bind with CBP(CREB binding protein) in vitro. To understand the interaction of fusion proteins and CBP, we studied the CBP protein in TC135 cells expressing the EWS-Fli-1 gene. We also studied the hypothesis that downregulation of fusion gene expression may induce susceptibility to apoptosis in Ewing's sarcoma cells. METHODS: For targeting fusion proteins, we reconstructed the antisense EWS-fli-l, EWS-erg and CBP genes in pcDNA3, and transfected these genes to Ewing's sarcoma cells showing high levels of expression for Ve3 and 5838 genes. These vectors were transfected to cells by the calcium phosphate method, and transformed cells were selected using G418. We measured DNA fragments for apoptosis using FACScan. We used crystal violet staining and MTT assay to evaluate cell viability, and Western blot analysis was used to assess CBP gene expression. RESULTS: Cells transfected with antisense fusion genes Ve3 and 5838 showed inhibition of fusion protein expression. These cells also showed decreased cell viability. Susceptibility to apoptosis was induced by treatment with chemotherapeutic agents at low concentrations. Antisense CBP- transfected cells showed loss of cell viability in O.l% and 0.5% serum. This loss of cell viability was similar to the response by antisense fusion protein-transfected cells treated with chemotherapeutic agents at low concentrations. CONCLUSION: Our results suggest that fusion proteins and CBP co-regulate apoptosis in Ewing's sarcoma cells. Antisense fusion gene therapy may be an useful adjunct in combining with chemotherapeutic regimens to downregulate the expression of fusion proteins in Ewing's sarcoma.
Apoptosis* ; Blotting, Western ; Calcium ; Cell Survival ; DNA ; Down-Regulation ; Gene Expression ; Genetic Therapy ; Gentian Violet ; Humans ; Sarcoma, Ewing* ; Transcription Factors

PURPOSE: We investigated compounds from food sources given to children that may induce the differentiation of small intestinal epithelial cells in order to signal pathways that induce the prolif eration and differentiation of small intestinal epithelial cells. METHODS: We analyzed small intestinal epithelial cell differentiation using in vitro IEC-6 cells model. The growth curve of IEC-6 cells was obtained by standard MTT assay. Alkaline phospha tase(ALP) activities were determined using the paranitrophenol colorimetric assay for the differ entiation of IEC-6 cells. We did ALP and Brdu double-staining of cultured IEC-6 cells to distin guish between differentiation and proliferation, and investigated compounds' potential for inducing differentiation of small intestinal epithelial cells and protein kinase signal pathway. RESULTS: The calcium ion was essential for the differentiation of IEC-6 cells. Retinol and retinoic acid induced the differentiation of IEC-6 cells. beta-LG stabilized and increased cell permeation of retinoic acid. IEC-6 cells showed 3 or 4 times more ALP activity with co-treatment of retinoic acid and beta-LG. BSA and OVA accelerated differentiation of IEC-6 cells in a similiar fashion to beta -LG. But, pepton and casein didn't. Heat destruction of beta-LG, BSA and OVA lead to loss in the ability of these compounds to induce cellular differentiation. The PKA signal pathway involved differentiation of IEC-6 cells. IEC-6 cells proliferation increased by the activation of PKC signal pathway and decreased differentiation by PKC signal pathway. CONCLUSION: Our results confirm that signal pathways are related to the proliferation and differ entiation of small intestinal epithelial cells and various compounds from food sources of childhood, such as beta-LG, BSA, OVA, and retinoic acid. These compounds appear to induce differentiation of small intestinal epithelial cells and may play a role in stimulating regeneration of epithelial cells after small intestinal mucosal injury.
Bromodeoxyuridine ; Calcium ; Caseins ; Child ; Epithelial Cells* ; Hot Temperature ; Humans ; Ovum ; Protein Kinases ; Regeneration ; Signal Transduction ; Tretinoin ; Vitamin A