Close linkage between X-linked ectodermal dysplasia and a cloned DNA sequence detecting a two allele restriction fragment length polymorphism in the region Xp11-q12

Summary EDA (ectodermal dysplasia, anhidrotic) is an X-linked recessive disorder characterized by hypohidrosis, hypoor anodontia, and hypotrichosis. A possible linkage between the gene for EDA and a number of restriction fragment length polymorphisms (RFLPs) spread over the X chromosome was investigated in two Danish families segregating EDA. No recombination between the gene for EDA and our probe pTAK8, which detects a two allele polymorphism in the region Xp11-q12, was found in nine informative meiotic events (seven of which are phase known), giving a maximal lod score of 2.41 at a recombination fraction of 0.00. This juxtacentromeric location of the gene for EDA agrees well with the linkage data obtained with the other markers used in this study.     

Model simulation of heat and water transport dynamics in an airway

Heat and water transport processes in the respiratory tract depend on environmental conditions, breathing patterns, and the physiological state of the respiratory system. To study these processes, we have developed a mathematical model of the dynamics of temperature and water vapor in the radial and axial directions of an idealized trachea. The model is expressed as two implicit finite-difference equations and solved using an alternating-direction algorithm. Using these equations, we simulated the effects of inspired gas temperature and humidity, velocity profile, and flow rate on heat and water transport between the gas and airway wall. Under inspired gas conditions of low temperature or high relative humidity, supersaturation occurs. Increasing either the velocity gradient at the wall or the flow rate increases the heat and water transport rates. However, these rates change by only 10 percent when the velocity gradient is doubled, and by about 35 percent when flow rate undergoes a two-fold change. The model can be used with in-vivo data from the trachea to test hypotheses concerning normal and abnormal heat and water transport.     

Enzymbestimmungen in Erythrocyten bei Kindern mit Down-Syndrom

Zusammenfassung Bei 20 Kleinkindern mit gesicherter freier Trisomie 21 wurden die Aktivitäten von 21 Enzymen des Energie- und des Glutathionstoffwechsels in isolierten Erythrocyten gemessen. Die Aktivitäten der Fructose-6-phosphatkinase, der Lactatdehydrgenase, der Glucose-6-phosphatdehydrogenase, der Glutathionreduktase (NADP) und der Glutamat-Oxalacetat-Transaminase wurden gegenüber gleichaltrigen Kontrollen signifikant erhöht gefunden. Lediglich die etwa 50%ige Steigerung der Fructose-6-phosphatkinase-Aktivität kann im Sinne eines Gen-Dosis-Effektes gedeutet werden.

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Enzyme activities in the erythrocytes of children with Down's syndrom

Summary The activities of 21 enzymes of the energy supplying and the glutathione metabolism were estimated in isolated erythrocytes from children with Down's syndrome aged from 1 to 5 1/2 years. All 20 patients were trisomic for chromosome 21. The following enzymes were estimated by means of optic assays: All glycolytic enzymes, two enzymes of the hexosephosphate shunt and two of the citric cycle, NADP- and NAD-dependent glutathione reductase, glutamic-oxaloacetic transaminase, adenylate kinase, and Mg²⁺-activated ATPase.The activities of five enzymes, i.e. phosphofructokinase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, glutathione reductase (NADP), and glutamic-oxaloacetic transaminase, were significantly elevated as compared to age-matched controls. The 50% increase of phosphofructokinase-activity is assumed to provide suggestive evidence for a controlling gene-locus on chromosome 21. The remaining findings are not considered to be directly related to the chromosomal abnormality.

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Direcktor: Prof. Dr. H.-R. Wiedemann

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Nach einem Vortrag auf der 10. Tagung der Gesellschaft für Anthropologie und Human-genetik, Königstein (Ts.), 22.–25. 10. 1967.

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Die Untersuchungen wurden durch die dankenswerte Unterstützung der Deutschen Forschungsgemeinschaft ermöglicht.     

PRODUCTION AND CHARACTERIZATION OF MULTIPLE-LAYERED POPULATIONS OF ANIMAL CELLS

Dense populations containing 129 x 10⁶ Jensen sarcoma, 134 x 10⁶ DON Chinese hamster, 28.9 x 10⁶ WI-38 human diploid, 61.8 x 10⁶ HEp-2 human carcinoma, and 67.4 x 10⁶ WISH human amnion cells were produced from dilute inocula, 0.85 to 5.33 x 10⁶, in 7 to 8 days in a perfusion system using replicate T-60 flasks. Perfusion rates as high as 560 ml medium/day/T-60 were required to maintain pH (to ca ±0.1 unit) and adequate nutrient supplies. The cell densities encountered are described by the term "monolayer equivalents" (M.E.), defined as number of cells per culture divided by number of cells in a monolayer. The M.E.'s for T-60 cultures containing unusually dense populations of 40 x 10⁶ WI-38 and 250 x 10⁶ DON cells (9-day perfusion) were 5 and 17, respectively, and numbers of cells in illustrations of stained cross-sections of membranes from these cultures were in excellent agreement. Threshold M.E.'s exist below which proliferation is the chief cellular activity and above which one or more cell functions may predominate even though proliferation persists. Cellular nutrition and metabolism may change with changes in M.E., as illustrated in different patterns of glutamic acid, proline, and glycine utilization or production in dense vs. dilute WI-38 cell populations. The results indicated that the role of contact inhibition phenomena in arresting cellular proliferation was diminished in perfusion system environments.     

Disinfection of Aerosolized Pathogenic Fungi on Laboratory Surfaces: II. Culture Phase

The effect of several fungicides on laboratory surfaces contaminated with the culture (spore) phase of aerosolized Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum was ascertained. The culture (spore) phase was more resistant to the action of the fungicides than was the tissue (yeast) phase. The addition of a wetting agent increased the efficiency of several fungicides. The time required for disinfection with a given concentration of fungicide, or the concentration required to disinfect within a given time, can be determined by interpolating the plotted graphs.     

Binding of the J 1 Adhesion Molecules to Extracellular Matrix Constituents

The J1 glycoproteins can be obtained in multiple forms in the soluble fraction of developing and adult mouse brain tissue. They are recovered as two forms of apparent molecular weights of 160,000 and 180,000 (J1-160) from adult mouse brain and as forms of apparent molecular weights of 200,000 and 220,000 (J1-220) from developing brain. J1-160 and J1-220 share common epitopes but are considered as separate entities, with J1-220 being immunochemically closely related if not identical to tenascin. Based on the observation that J1 immunoreactivity appears on basement membrane and interstitial collagens after denervation of the neuromuscular junction in adult rodents, we became interested in investigating the binding properties of J1 glycoproteins to extracellular matrix constituents in vitro. Both J1-160 and J1-220 bound to collagens type I-VI and IX but not to laminin, fibronectin, bovine serum albumin, or gelatin under hypotonic buffer conditions. Under isotonic buffer conditions, J1-220 bound to all collagen types, whereas J1-160 bound only to collagen types V and VI with values that could be examined by Scatchard analysis. Binding of J1-220 to collagens displayed two binding constants (KD) between 1.5 and 4.4 X 10(-9) and 1.8 and 5.5 X 10(-8) M, respectively, under hypotonic buffer conditions and a single KD of 2.1-8.0 X 10(-8) M under isotonic buffer conditions. Binding of J1-160 to collagens had an apparent KD of 1.9-8.0 X 10(-9) M under hypotonic buffer conditions. Under isotonic buffer conditions, binding constants of J1-160 to collagen types V and VI were approximately 2 X 10(-8) M. Binding of J1-220 to collagen type I could be inhibited by J1-220, J1-160, and collagen type VI but not by fibronectin or gelatin. Conversely, binding of J1-160 was inhibited by J1-220, J1-160, and collagen type VI (in order of decreasing efficacy of competition). J1-160 and J1-220 were retained on a heparin-agarose column and eluted in a salt gradient at approximately 0.5 M NaCl. The formation of the J1-heparin complexes was inhibited 100-fold more efficiently by heparin than by chondroitin sulfate. These experiments show that J1 glycoproteins resemble in many respects the extracellular matrix constituents fibronectin, laminin, vitronectin, and von Willebrand factor.     

Multipoint linkage analysis in Menkes disease.

Linkage analyses were performed in 11 families with X-linked Menkes disease. In each family more than one affected patient had been diagnosed. Forty informative meioses were tested using 11 polymorphic DNA markers. From two-point linkage analyses high lod scores are seen for DXS146 (pTAK-8; maximal lod score 3.16 at recombination fraction [theta] = .0), for DXS1 (p-8; maximal lod score 3.44 at theta = .0), for PGK1 (maximal lod score 2.48 at theta = .0), and for DXS3 (p19-2; maximal lod score 2.90 at theta = .0). This indicates linkage to the pericentromeric region. Multilocus linkage analyses of the same data revealed a peak for the location score between DXS146(pTAK-8) and DXYS1X(pDP34). The most likely location is between DXS159 (cpX289) and DXYS1X(pDP34). Odds for this location relative to the second-best-supported region, between DXS146(pTAK-8) and DXS159 (cpX289), are better than 74:1. Visualization of individual recombinant X chromosomes in two of the Menkes families showed the Menkes locus to be situated between DXS159(cpX289) and DXS94(pXG-12). Combination of the present results with the reported absence of Menkes symptoms in male patients with deletions in Xq21 leads to the conclusion that the Menkes locus is proximal to DXSY1X(pDP34) and located in the region Xq12 to Xq13.3.