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J Kunz

Publications and source records attributed to J Kunz.

At least 91 records · Page 5Linked to original sources
PubMed1994

Regional localization of 725 human chromosome 7-specific yeast artificial chromosome clones.

Toward the construction of a complete physical map of human chromosome 7, we have localized 725 YAC clones to cytogenetically defined regions using fluorescence in situ hybridization (FISH) and by screening with DNA markers of known chromosomal locations. These chromosome 7-specific YAC clones are part of a library constructed with DNA isolated from monochromosomal 7 human-hamster somatic cell hybrid lines. The FISH mapping for 575 clones was accomplished by using "Alu-PCR" amplified YAC DNA against metaphase chromosome spreads made from a monochromosomal 7 human-mouse somatic cell hybrid line. Hybridization- or PCR-based screening of previously mapped DNA markers was performed for the mapping of 221 YAC clones. There was excellent correlation between the map locations obtained for the 71 YACs localized with both methods. All of the regionally localized YAC clones are valuable reagents for mapping and identification of disease genes on human chromosome 7.

Animals↗
PubMed1994

An optimized method for routine HLA-B27 screening using flow cytometry.

Flow cytometry and monoclonal antibodies are promising tools for HLA-antigen detection. Previous approaches have been hampered by the lack of a carefully standardized system for calibration and sample analysis. A new system for HLA-B27 screening was developed using a FACScan flow cytometer, software for automated calibration and analysis, calibration beads, and the anti-HLA-B27-FITC/anti-Leu4-PE (CD3) monoclonal antibodies. The median fluorescence channel result for the HLA-B27-FITC signal of CD3+ T lymphocytes is compared to a decision marker. Values lower than this threshold are read as HLA-B27 negative and those above are recommended for retesting with the classic microcytotoxicity assay on the presumption of HLA-B27 positivity. The anti-HLA-B27 antibody reacts with all six HLA-B27 subtypes and shows a weaker binding to HLA-B7. The screening test results were compared with those from the microcytotoxicity assay for HLA-typing in studies involving several European centers. The observed sensitivity was 100% (95% Cl:98.6-100) and the specificity was 97.4% (95% Cl: 96.4-98.3). Other performance studies verified the reproducibility and reliability of results obtained with the screening system.

Anticoagulants↗
PubMed1994

The 140-kDa protein of blood-brain barrier-associated pericytes is identical to aminopeptidase N.

Although ample evidence has been accumulated on the structure and functional role of endothelial cells in blood-brain barrier mechanisms, little is known about the contribution that cerebral pericytes provide to this phenomenon. We have reported recently on a monoclonal antibody specific for cerebral pericytes at blood-brain barrier sites. To confirm the pericytic localization of this antigen, and in order to elucidate its biochemical identity, we have performed immunocytochemical, biochemical, and molecular biological studies. By immunocytochemistry on the light microscopic as well as electron microscopic level, we provide definite evidence that the 140-kDa antigen recognized by this monoclonal antibody is confined to cerebral pericytes, whereas endothelial cells are devoid of this antigen. N-Terminal sequencing of the corresponding immunocrossreacting renal protein revealed that the protein detected by the monoclonal antibody is identical to aminopeptidase N. By means of the reverse transcriptase-polymerase chain reaction, the identity of the 140-kDa antigen as aminopeptidase N could also be verified for cerebral microvascular cells. Cerebral pericytic aminopeptidase N may be involved in neurotransmitter (enkephalin) metabolism at the blood-brain interface. By taking into account that brain pericytes have been found to express further plasma membrane-bound enzymes, these results strongly suggest the contribution of cerebral pericytes in the metabolic concert of the homeostatic balance regulated by the blood-brain barrier.

Amino Acid Sequence↗
PubMed1994

TOR1 and TOR2 are structurally and functionally similar but not identical phosphatidylinositol kinase homologues in yeast.

The Saccharomyces cerevisiae genes TOR1 and TOR2 were originally identified by mutations that confer resistance to the immunosuppressant rapamycin. TOR2 was previously shown to encode an essential 282-kDa phosphatidylinositol kinase (PI kinase) homologue. The TOR1 gene product is also a large (281 kDa) PI kinase homologue, with 67% identity to TOR2. TOR1 is not essential, but a TOR1 TOR2 double disruption uniquely confers a cell cycle (G1) arrest as does exposure to rapamycin; disruption of TOR2 alone is lethal but does not cause a cell cycle arrest. TOR1-TOR2 and TOR2-TOR1 hybrids indicate that carboxy-terminal domains of TOR1 and TOR2 containing a lipid kinase sequence motif are interchangeable and therefore functionally equivalent; the other portions of TOR1 and TOR2 are not interchangeable. The TOR1-1 and TOR2-1 mutations, which confer rapamycin resistance, alter the same potential protein kinase C site in the respective protein's lipid kinase domain. Thus, TOR1 and TOR2 are likely similar but not identical, rapamycin-sensitive PI kinases possibly regulated by phosphorylation. TOR1 and TOR2 may be components of a novel signal transduction pathway controlling progression through G1.

1-Phosphatidylinositol 4-Kinase↗
PubMed1993

Target of rapamycin in yeast, TOR2, is an essential phosphatidylinositol kinase homolog required for G1 progression.

The yeast TOR2 gene encodes an essential 282 kd phosphatidylinositol (PI) 3-kinase homolog. TOR2 is related to the catalytic subunit of bovine PI 3-kinase and to yeast VPS34, a vacuolar sorting protein also shown to have PI 3-kinase activity. The immunosuppressant rapamycin most likely acts by inhibiting PI kinase activity because TOR2 mutations confer resistance to rapamycin and because a TOR1 TOR2 double disruption (TOR1 is a nonessential TOR2 homolog) confers G1 arrest, as does rapamycin. Our results further suggest that 3-phosphorylated phosphoinositides, whose physiological significance has not been determined, are an important signal in cell cycle activation. In yeast, this signal may act in a signal transduction pathway similar to the interleukin-2 signal transduction pathway in T cells.

Amino Acid Sequence↗
PubMed1993

All known human H1 histone genes except the H1(0) gene are clustered on chromosome 6.

PCR analysis of chromosomal DNA from a panel of human-rodent somatic cell hybrids revealed that the five human H1 histone genes H1.1 to H1.5 and the gene encoding the testis-specific H1t subtype, all of which form clusters with core histone genes, are located on chromosome 6. The H1(0) subtype, which is not neighbored by core histone genes, maps to chromosome 22. Fluorescence in situ hybridization with human metaphase chromosomes and PCR analysis of somatic cell hybrid DNA carrying only fragments of chromosome 6 revealed the region 6p21.1 to 6p22.2 as the histone gene cluster region.

Animals↗
PubMed1993

Chromosomal mapping of the human gene for the tricyclic antidepressant-sensitive noradrenaline transporter.

The neuronal Na(+)- and Cl(-)-dependent transporter for the neurotransmitter noradrenaline (NA) is the primary target for the tricyclic antidepressant desipramine. The NA-transporter belongs to a new gene family of structurally related Na(+)- and Cl(-)-dependent neurotransmitter transporters. In this study, the chromosomal localization for the gene encoding the human NA-transporter (h-NAT) was determined. By hybridization of a panel of somatic cell hybrids and by fluorescent in situ hybridization to metaphase chromosomes, the hNAT gene was localized on chromosome 16q12.2. In addition, evidence is presented for an intron-exon structure of the gene.

Animals↗
PubMed1993

Overexpression of HER2/neu oncogene in human pancreatic carcinoma.

The HER2/neu oncogene encodes a transmembrane protein that possesses intrinsic tyrosine kinase activity. Its overexpression has been associated with the malignant phenotype. In this study we examined HER2/neu expression in the normal and cancerous human pancreas. In the normal pancreas HER2/neu immunostaining was observed in acinar and ductal cells. HER2/neu immunoreactivity was expressed in 34 of 76 (45%) pancreatic carcinomas. There was a significant correlation between tumors with well-differentiated histology and HER2/neu expression. Northern blot analysis demonstrated HER2/neu mRNA expression in the normal pancreas and in situ hybridization confirmed its distribution in both acinar and ductal cells. In cancer tissues Northern blot analysis indicated that HER2/neu mRNA levels were elevated in 13 of 25 (52%) of the tumors in comparison with the normal tissues. In addition, in situ hybridization demonstrated a strong but heterogenous distribution of mRNA grains in these tumors. Southern blot analysis did not demonstrate HER2/neu gene amplification in any of the tumors. These data indicate that the HER2/neu protein is synthesized in the normal exocrine pancreas and is frequently overexpressed in well-differentiated adenocarcinomas of the pancreas as a result of increased HER2/neu mRNA levels.

Adult↗
PubMed1993

Cyclosporin A, FK506 and rapamycin: more than just immunosuppression.

The mechanisms of action of the immunosuppressive drugs cyclosporin A (CsA), FK506 and rapamycin are strikingly conserved from yeast to human T cells. Recent results obtained with yeast corroborate calcineurin as the target of CsA-cyclophilin and FK506-FKBP complexes, and reveal a phosphatidylinositol 3-kinase homologue as the target of the rapamycin-FKBP complex.

Amino Acid Isomerases↗