Search

Objective: The Jarvik 2000 (Jarvik Heart, Inc, New York, NY) is a thumb-sized high-speed impeller pump that is used as a ventricular assist device in patients with terminal heart failure. Because the Jarvik 2000 is designed for long-term use, it is a central question whether the mechanical forces inside the pump affect blood components. This study evaluated the potential association of the high rotational speed of the Jarvik 2000 with platelet activation, which may result in thromboembolic events., Methods: The study group comprised patients with terminal heart failure who were supported with the Jarvik 2000. All were men and received 100 mg aspirin daily. In 8 patients, soluble platelet activation markers (P-selectin and sCD40L), platelet counts, and hemolysis markers (haptoglobin and lactate dehydrogenase levels) were determined. In 5 patients, P-selectin expression and platelet receptor glycoprotein IIb/IIIa activation were determined with flow cytometry and compared with a control group of 5 healthy men. Platelet activation was measured at various rotational device speeds., Results: After Jarvik 2000 implantation, increased hemolysis was observed, but platelet activation markers and platelet counts were not affected. Increased rotational speed (8000 to 12,000 rpm) of the device also did not result in increased platelet activation., Conclusion: The Jarvik 2000 was not associated with detectable platelet activation, despite high rotational impeller speeds.

Objectives: In ageing population, both Alzheimer's disease (AD) and depression are common. Significant depressive symptoms are often co-morbid with cognitive impairment and dementia. In this study, we attempted to find various factors and markers for both AD and depression in a longitudinal cohort, the Vienna-Transdanube-Aging (VITA)-study., Methods: The VITA-Study consisted of 305 healthy subjects, 174 subjects with depression only, 55 subjects diagnosed with AD only and 72 subjects with depression as well as AD. Associations between AD and/or depression to gene polymorphisms APO E (epsilon4), choline acetyltransferase (ChAT) 4G to A, serotonin-transporter gene promoter-length, dopamine-D4-receptor, ciliary-neurotrophic-factor-null mutation and brain-derived neurotrophic factor (C270T) and to various known factors were analyzed., Results: AD and depression were significant associated. Significant risk factors found for AD were low education, low folic acid and depressive-symptoms, while for depression were low education and higher nonsteroidal anti-inflammatory drugs (NSAID) consume. Moreover, the ChAT polymorphism associated significant to depression. Gender, education, and ChAT significantly associated with the combination AD and/or depression., Conclusion: Such studies must be conducted cautiously, as co-morbidities and gene-environmental-social influences may sway the results dramatically. We found in the VITA-study significant association between depression and AD and between ChAT polymorphism and depression.

Depending on the type and localisation, nonunions of tibial fractures will occur in 10-40% of cases. Bone morphogenetic protein 7 (BMP-7; Osigraft), a recombinant bone growth factor, can be implanted locally as an alternative to autologous bone grafting. The objective of our study was to compare the efficiency of the two procedures. From January 1995 to December 2002, 82 patients (group 1) with delayed union of a tibial fracture received autologous bone grafting as their first procedure. To compare their results with the efficiency of BMP-7, between May 2002 and June 2005 we followed up on 26 patients (group 2) who had local implantation of BMP-7 after having had, on average, four surgical procedures. Healing was considered successful if x-rays showed bony consolidation and if no further procedure was necessary. Group 1 had no signs of consolidation in 24 cases (28%), whereas group 2 had only two (8%) such patients (p=0.025). The BMP-7 group showed a significantly higher success rate compared with patients with autologous bone grafting, despite the fact that the BMP-7 group contained more complicated cases.

HLA-E is a ligand for the immune-inhibitory NKG2A receptor expressed on NK and T cells. To investigate HLA-E expression and immune cell infiltration in human astrocytic tumors in vivo, we analyzed normal CNS controls and astrocytomas of all WHO grades by immunohistochemistry. Both, CD8(+) immune cell infiltration and HLA-E expression were significantly higher in astrocytic tumors than in normal brain. Further, HLA-E expression levels and immune cell infiltration were significantly correlated in WHO grade IV glioblastomas. Thus, HLA-E overexpression in glioblastomas may be triggered by T and NK cell infiltration.

Purpose: To evaluate acute changes in atrial and ventricular parameters by the use of cardiac magnetic resonance imaging (MRI) in patients with percutaneous transcatheter atrial septal defects (ASD) closure., Materials and Methods: The study included 14 patients (six males and eight females, 45 +/- 18 years) with congenital ASD. Cardiac MRI (1.5T Philips Intera CV) was performed before and within 24 hours after transcatheter ASD closure. Right atrial (RA) and left atrial (LA) dimensions, as well as right (RV) and left (LV) ventricular end-diastolic (ED) volumes were determined. Atrial size was assessed by planimetry of the maximum RA and LA areas in a standard four-chamber view, and ventricular volumes were calculated according to a modified Simpson's rule in short-axis views., Results: The mean RA decreased significantly from 27.6 +/- 6.4 cm(2) before closure to 24.4 +/- 5.6 cm(2) after the procedure (P = 0.0018), whereas the LA area did not change (24.1 +/- 4.7 cm(2) vs. 23.8 +/- 5.2 cm(2), P = 0.76). The RV volumes, volume index, and ejection fraction (EF) decreased significantly from 229 +/- 64 mL to 181 +/- 43 mL (P < 0.001, average reduction = 19% +/- 15%), from 126.0 +/- 37.2 mL/m(2) to 96.6 +/- 28.6 mL/m(2) (P < 0.0001) and from 64 +/- 5% to 58% +/- 7% (P = 0.01), respectively. The LV volumes and volume index remained unchanged (114 +/- 25 mL vs. 118 +/- 22 mL, P = 0.18, 63.5 +/- 13.5 mL/m(2) vs. 63.0 +/- 17.4 mL/m(2), P = 0.83). Left-right shunting decreased from 40% +/- 15% to 9% +/- 15% (P < 0.001)., Conclusion: Cardiac MRI can reveal detailed information on acute changes in shunt fraction and ventricular dimensions after ASD closure. ASD closure by percutaneous transcatheter device implantation results within 24 hours in a significant reduction of shunt fraction, RA and RV sizes, and RV function, whereas LA and LV dimensions remain unchanged., ((c) 2007 Wiley-Liss, Inc.)

Background: The site of effect for the selective serotonin reuptake inhibitors (SSRIs) is the serotonin transporter (5-HTT), which is extensively investigated for its involvement in depressive symptoms. The 5-HTT gene exhibits a 5'-promoter-based length polymorphism (5-HTTLPR) that affects the transcription efficiency and activity, known as short (S) and long (L) alleles. We studied the association of this polymorphism in old age and depression in the Vienna Transdanube Aging (VITA) study, excluding subjects with dementia., Method: We used retrospective data from the baseline of the VITA study, which is a cohort study of all inhabitants of a geographical area aged 75 years (N = 544). Depression was diagnosed and classified strictly according to the DSM-IV. To eliminate dementia effects, we excluded subjects with a Clinical Dementia Rating higher than or equal to 1 and/or a Mini-Mental State Examination score lower than 24. Genotyping for the 5-HTTLPR L/S allele was conducted using polymerase chain reaction methodology., Results: We found significantly higher SS genotype frequency in all subjects with past/ present depression compared to controls (trend test, p = .01). The SS genotype frequency was especially high in subjects with onset of depression before age 65. No correlations were found between genotypes/S allele carriers and actual Hamilton Rating Scale for Depression, Short-Geriatric Depression Scale, and anxiety scale scores., Conclusions: These observations of higher frequency of the 5-HTTLPR S allele in subjects with past/present depression fit with previous findings and point to the important role of 5-HTT in depression.

Purpose: To assess the geometry and area of the left ventricular outflow tract (LVOT) in non-stenotic and stenotic aortic valves and to determine the aortic valve area (AVA) in non-stenotic valves by magnetic resonance imaging (MRI) using a modified continuity equation., Methods: Twenty patients (10 male, mean age 54.8 +/- 15 years) without known aortic valve disease and 10 patients (7 male, mean age 65.1 +/- 14 years) with moderate to severe aortic stenosis were included in this study. MRI was performed using a 1.5 T scanner (Philips Intera CV). AVA was assessed by planimetry on high quality SSFP cine sequences and used as reference standard. LVOT area was defined by calculating a circular area using the LVOT diameter from the 3 chamber view (3CV) and by planimetry. Peak flow velocity was assessed in the LVOT and the proximal aorta. AVA was calculated by a modified Gorlin equation, the continuity equation and a modified continuity equation using the planimetric LVOT area., Results: Planimetric AVA ranged from 2.9 to 6.4 cm2 in patients with non-stenotic and from 0.3 to 1.3 cm2 with stenotic valves, LVOT area from 3.4 to 6.1 cm2 and from 2.6 to 6.5 cm2, respectively. The LVOT area based on the LVOT diameter derived from the 3CV was significantly underestimated in comparison to planimetry in non-stenotic and stenotic aortic valves (3.3 +/- 0.7 vs. 4.7 +/- 1.0 cm2, p < 0.0001; mean difference 1.1 +/- 0.12 cm2, CI 0.86-1.36 and 3.7 +/- 1.2 vs. 4.7 +/- 1.5 cm2, p < 0.05; mean difference 1.0 +/- 1.0 cm2, CI 0.24-1.71). The Gorlin formula showed a poor agreement with planimetry, whereas continuity equation and the modified continuity equation revealed a very good agreement. Planimetry of the LVOT displayed an elliptic shape of the LVOT in all patients with the minimum diameter perpendicular to the 3CV, which was the reason for the above mentioned underestimation., Conclusion: The LVOT area calculated from the 3CV-LVOT diameter underestimates the LVOT area compared to planimetry due to an elliptic shape of the LVOT in patients with non-stenotic as well as with stenotic aortic valves. The modified Gorlin equation proved to be less useful to assess AVA in non-stenotic valves, whereas the continuity equation and a modified continuity equation displayed a very good agreement with planimetric area measurements.

Objective: Within the multiprofessional diagnostic in the psychiatric treatment for children an age-group of in-patients was examined using the coordination test for children (KTK) with regard to physical ability., Methods: The ascertained results and classifications of the motor skills were related to ICD 10 diagnosis on level 1 (clinical psychiatric syndrom) and level 2 (circumscribed disturbance of the development) as well as to issues of the anamnese- and report-documentation in order to ascertain correlations between child psychiatry disorder and motor development., Results: More than 40 % of the children examined showed motor deficiency. Moreover, it was evident that girls attained significantly lower KTK-results than boys. The statistical results did not show any distinct effects to "diagnosis" and "age"., Conclusions: The high quota of obviously impaired and disturbed children reflects the general decline of motor skills and does not seem to be attributed to the demand for psychiatric treatment for children and youth.

For testing with sodium lauryl sulphate (SLS), measurements of transepidermal water loss (TEWL) and cutaneous blood flow with laser Doppler (LD) are considered to be the most reliable methods. The aim of this study was to determine which method of measurement should be preferred when conducting SLS testing under varying conditions. Patch testing with SLS at different concentrations and exposure times was performed. TEWL values were compared with those of LD. TEWL values showed distinct changes at low SLS concentrations and short application periods. By contrast, higher SLS concentrations were necessary to increase LD values. Short application of patches changed TEWL rather than LD values. When evaluating SLS patch testing by bioengineering methods, TEWL measurement appears to be more suitable for a test procedure that provokes mild skin reactions (SLS concentration <1%), whereas LD measurement is more appropriate to evaluate pronounced skin reactions (SLS concentration >or=1%).

Background: It is well known that the degree of skin reaction to an irritant depends on its concentration and exposure time., Objectives: To determine the interrelationship between the concentration of sodium lauryl sulphate (SLS) and exposure time in both weak (subclinical) and severe reactions., Methods: Patch testing with SLS was performed at different concentrations (0.125%, 0.25%, 0.5%, 1.0% and 2.0%) and with different exposure times (3, 6, 12, 24 and 48 h). Evaluation was conducted by measurement of transepidermal water loss and by laser-Doppler flowmetry both 30 min and 24 h after patch removal., Results: We found more reliable and constant skin reactions 24 h after patch removal, and a higher correlation between SLS concentration and skin reaction., Conclusions: We conclude that the concentration of SLS influences the test outcome to a larger degree than the exposure time. We present formulae by which the outcome of SLS patch testing at various SLS concentrations ranging from 0.125% to 2% and any exposure time between 3 and 24 h can be estimated.

Mutations in the human gene Jagged1 (JAG1) localized in 20p12 have been recently identified as causal for the anomalies found in patients with Alagille syndrome (AGS). This gene encodes a ligand for the Notch1 transmembrane receptor, which plays a key role in cell-to-cell signaling during differentiation and is conserved from C. elegans to human. We report a paracentric inversion (PAI) of chromosome 20p12.2p13 in an individual with AGS who also had alpha-1-antitrypsin deficiency. To our knowledge, this is the first published case of PAI involving the short arm of chromosome 20. Using FISH, fiberFISH, and molecular studies with a approximately 40 kb cosmid clone encompassing the entire 36 kb JAG1 gene, we demonstrate that the gene was disrupted by the inversion breakpoint between exons 5 and 6. An unusual association between two most common causes of chronic liver disease in childhood, AGS and alpha-1-antitrypsin deficiency, as well as their influence on the proband's abnormal phenotype are discussed., (Copyright 2001 Wiley-Liss, Inc.)

The flow cytometric detection of aberrant antigen expression is one method proposed for the quantification of minimal residual disease (MRD) in acute leukemias. The present study was designed to investigate the stability of the aberrant antigen expression at relapse or at treatment failure of initial chemotherapy. For this purpose, multiparameter immunophenotyping with a panel of 15 monoclonal antibodies was used at diagnosis as well as at relapse (43 patients with overall 65 aberrations) and at treatment failure (35 patients with overall 66 aberrations). There was a significant decrease in the percentage of the initially described aberrant antigen expression on leukemia blasts at relapse (P = 0.001; n = 65) as well as at treatment failure (P = 0.0001; n = 66) considering all aberrations in the whole leukemia population. Concerning only patients with acute myelogenous leukemia (AML), significant decreases in the aberrant expression could be detected at relapse (P = 0.031; n = 42) and at treatment failure (P = 0.0001; n = 52). The changes in patients with acute lymphoblastic leukemia (ALL) were significant only at relapse (P = 0.006; n = 23). Initially, the most informative aberration was not detectable in four patients at relapse and in seven patients at treatment failure. A decrease of under 50% of the initial value was observed in another 8 patients at relapse and in 10 patients at treatment failure. In further studies assessing the detection of aberrant antigen expression for MRD, quantification of the relapses should be explicitly analyzed regarding the persistence of the initially described aberrant antigen expression., (Copyright 2000 Wiley-Liss, Inc.)

In vascular smooth muscle, openers of ATP-dependent potassium channels (K(ATP) channels), such as P1075 (N-cyano-N'-(1,1-dimethylpropyl)-N"-3-pyridylguanidine), produce relaxation. In this study we have investigated the effects of thiol-modifying agents on the binding of P1075 and on the 86Rb+ efflux stimulating and vasorelaxant effects of the opener in rat aortic rings. The increase in 86Rb+ efflux induced by P1075 was taken as a qualitative measure of K+ channel opening. The hydrophilic SH-group-oxidizing substance, thimerosal (1 to 100 microM), abolished specific binding of [3H]-P1075 with an IC50 value of 7.6+/-1.2 microM; at 30 microM, the half time for inhibition was 38 min. Two other thioloxidizing agents, PMB (4-hydroxy-mercuribenzoic acid) and DTBNP (2,2'-dithio-bis(5-nitropyridine)), inhibited binding up to 86% and 44%, respectively. The disulphide bond reducing substance, DTT (1,4-dithiothreitol, 0.1 to 1 mM), reduced [3H]-P1075 binding by up to 20% and partially reversed the inhibitory effect of thimerosal. In 86Rb+ efflux experiments, thimerosal (3 to 100 microM) concentration-dependently increased basal efflux but inhibited P1075-stimulated tracer efflux with an IC50 value of 7+/-1 microM. The inhibitory effect occurred with a half-time of approximately 8 min and was essentially reversed by DTT. In rings precontracted with noradrenaline, thimerosal inhibited the vasorelaxant effect in a noncompetitive manner, shifting the concentration-relaxation curves to the right and reducing maximum relaxation. The data show that oxidation of thiol groups interferes with the binding of the K(ATP) channel opener, P1075; concomitantly, the 86Rb+ efflux stimulating and the vasorelaxant effects are inhibited. Reduction of disulphide bonds by DTT has only minor effects on the action of P1075. Collectively, the results suggest that intact thiol groups are essential for the functioning of the K(ATP) channel in rat aorta. The different kinetics governing the inhibition of opener binding and of opener-stimulated 86Rb+ efflux suggest that the SH-groups involved in the two processes differ in their accessibility to thimerosal and/or in their reactivity.

In rat aortic rings, P1075, an opener of ATP-dependent potassium channels (K(ATP) channels), produces relaxation and 86Rb+ efflux from preloaded tissues; the increase in 86Rb+ efflux qualitatively reflects K(ATP) channel opening. In this study we have investigated the effects of protein kinase C modulation on the 86Rb+ efflux stimulating, the vasorelaxant and the binding properties of P1075. Phorbol 12,13-dibutyrate (PDBu), a direct activator of protein kinase C, inhibited the 86Rb+ efflux produced by P1075 with an IC50 value of 20+/-2 nM. Phorbol 12-myristate 13-acetate (PMA), another stimulator of protein kinase C, was 150 times weaker in this respect whereas 4alpha-PDBu, the inactive stereoisomer of PDBu, was ineffective. Staurosporine (300 nM), an inhibitor of protein kinase C, induced a small but significant increase of P1075-induced tracer efflux and partially reversed the inhibitory effect of PDBu on P1075-stimulated tracer efflux. The vasorelaxant effect of P1075 was inhibited only to a moderate degree by PDBu at concentrations which inhibited P1075-induced 86Rb+ efflux to >90%; however, in the presence of PDBu, the relaxation kinetics of P1075 were increasingly slowed. The vasorelaxant effect of P1075 in the presence of PDBu was still sensitive to inhibition by glibenclamide (100 nM), the standard inhibitor of the K(ATP) channel openers. Specific binding of [3H]-P1075 to rat aortic rings was unaffected by PDBu and PMA even in the micromolar concentration range. The data show that stimulation of protein kinase C inhibits the K+ channel opening effect of P1075 in rat aorta and suggest that protein kinase C may exert a weak tonic inhibition on the K(ATP) channels in this vessel under quasiphysiological conditions. At concentrations of PDBu which essentially abolished P1075-induced tracer efflux, the glibenclamide-sensitive vasorelaxant effect of P1075 was slowed down but not prevented; this supports earlier suggestions that K+ channel openers are also able to relax smooth muscle cells by a mechanism independent of K(ATP) channel opening.

The effect of activators of protein kinase A on membrane K+ permeability and the interaction of these compounds with cromakalim, an opener of ATP-sensitive K+ channels (K(ATP) channels), were investigated. Membrane K+ permeability was assessed by measuring 86Rb+ efflux from rings of rat aorta. Forskolin, an activator of adenylate cyclase, and isobutylmethylxanthine (IBMX), a nonselective phosphodiesterase inhibitor, induced small, concentration-dependent increases in tracer efflux up to 20-40% over the basal level. The effect of forskolin was abolished by the K+ channel blocker tedisamil (1 microM) and partially inhibited by glibenclamide (1 microM), a relatively selective blocker of K(ATP) channels. Further studies were conducted in the presence of 35 mM KCI in the bath in order to increase the size of the 86Rb+ efflux stimulated by forskolin and IBMX. At high concentrations, these compounds produced a biphasic effect with a peak increase being followed by a lower plateau value. Glibenclamide inhibited the 86Rb+ efflux response to forskolin and IBMX by 50-80%. The K+ channel blockers tedisamil (1 microM), Ba2+ (1 mM) and tetraethylammonium (10 mM) also reduced the peak response to forskolin by about 50% and abolished or greatly inhibited the plateau response. In addition to the small effect on basal 86Rb+ efflux, forskolin (0.3 microM) increased cromakalim-induced 86Rb+ efflux 3.4 times. At higher concentrations, however, a concentration-dependent inhibition was observed with an IC50 value of 7.6 +/- 0.4 microM. 1,9-dideoxyforskolin, which does not increase cAMP, increased neither basal nor cromakalim-induced 86Rb+ efflux; however, it inhibited cromakalim-stimulated tracer efflux with an IC50 value of 22 +/- 2 microM. It is concluded that forskolin and IBMX, probably by increasing intracellular cAMP levels, induce a 86Rb+ efflux from rat aorta, the major part of which is glibenclamide-sensitive and may pass through K(ATP) channels. In addition, low concentrations of forskolin greatly facilitate the K(ATP) channel opening effect of cromakalim whereas high concentrations block the channel; this blocking effect of forskolin is unrelated to the cAMP elevating action.

1. The binding of the sulphonylurea [3H]-glibenclamide, a blocker of adenosine 5'-triphosphate (ATP)-sensitive K+ channels (KATP channels), was studied in endothelium-denuded rings from rat aorta. 2. [3H]-glibenclamide labelled two classes of binding sites with KD values of 20 +/- 5 nM and 32 +/- 1 microM. The high affinity component, which comprised 17% of total binding at 1 nM [3H]-glibenclamide, had an estimated binding capacity of 150 fmol mg-1 wet weight. 3. Other sulphonylureas such as glipizide and glibornuride and the sulphonylurea-related carboxylate, AZ-DF 265, inhibited high affinity [3H]-glibenclamide binding with the potencies expected from their K+ channel activity. At very high concentrations, AZ-DF 265 and glipizide started to interact also with the low affinity component of [3H]-glibenclamide binding. 4. Openers of the ATP-sensitive K+ channel belonging to different structural groups inhibited only the high affinity [3H]-glibenclamide binding; the potencies in this assay were similar to those obtained in functional (i.e. vasorelaxation) studies. 5. High affinity [3H]-glibenclamide binding was abolished by prolonged hypoxia combined with metabolic inhibition. 6. The data indicate that the high affinity component of [3H]-glibenclamide binding mediates the block of the KATP channel by the sulphonylureas in rat aorta; hence, it represents the sulphonylurea receptor in this vessel. The pharmacological properties of this binding site resemble those of the binding site for the openers of the KATP channel; present evidence suggests that these two classes of sites are negatively allosterically coupled.

The kidney is endowed with ATP-sensitive K+ channels (KATP channels) both at the vascular and at the epithelial level. In this study we have characterized the binding of the sulphonylurea glibenclamide, the most widely used blocker of KATP channels, in rat isolated glomeruli. In metabolically intact glomeruli, 3H-glibenclamide labelled two different binding components with affinities of 47 +/- 12 nM and 10 +/- 1 microM and estimated binding capacities of 1.2 +/- 0.1 and 501 +/- 11 pmol/mg protein, respectively. 3H-glibenclamide binding was inhibited differentially by other sulphonylureas (tolbutamide, glibornuride, gliquidone and glipizide) and benzoic acid analogues such as meglitinide, AZ-DF 265 and UL-DF 9. Sulphonylureas interacted with the high affinity component and, in some cases, also with the low affinity component whereas the benzoic acid derivatives inhibited exclusively low affinity glibenclamide binding. Severe metabolic stress affected both components of glibenclamide binding by shifting high affinity binding to the right and reducing the capacity of the low affinity component. Disruption of the cytoskeletal actin filaments by cytochalasin B and D mimicked the effect of metabolic stress on the high affinity component but left the low affinity component unchanged. In crude membranes, the affinity of the first component was again reduced and a major loss of the low affinity sites was observed. The data show that the two binding components of glibenclamide binding in rat isolated glomeruli have very different properties. The high affinity component is not recognized by the benzoic acid derivatives; its affinity is modulated by cell metabolism and the actin component of the cytoskeleton. The low affinity sites are, in their majority, cytosolic. The function and cellular localization of the high affinity sites are under further study.

Karyotypes with an interstitial deletion and a marker chromosome formed from the deleted segment are rare. We identified such a rearrangement in a newborn infant, who presented with macrocephaly, asymmetric square skull, minor facial anomalies, omphalocele, inguinal hernias, hypospadias, and club feet. The karyotype 46,XY,del(5) (pter --> p13::cen --> qter)/47,XY,+dicr(5)(:p13 --> cen::p13 --> cen), del(5)(pter --> p13::cen --> qter) was identified by banding studies and FISH analysis in the peripheral lymphocytes. One breakpoint on the del(5) maps distal to GDNF, and FISH analysis using an alpha-satellite probe suggests that the proximal breakpoint maps within the centromere. The dicentric r(5) consists of two copies of the segment deleted in the del(5), resulting in trisomy of proximal 5p (5p13-cen). The phenotype of the propositus is compared with other trisomy 5p cases and possible mechanisms for the generation of this unique chromosomal rearrangement are discussed.

The human amyloid precursor-like protein APLP2 is a highly conserved homologue of a sequence-specific DNA-binding mouse protein with a predicted function in the cell cycle. Somatic cell hybrids segregating human chromosomes were used to assign the APLP2 gene to chromosome 11. Fluorescence in situ hybridization confirmed this assignment and further localized the gene to q23-q25.

Bone morphogenetic protein 2A (BMP2A), a member of the decapentaplegic-Vg-related family, belongs to the transforming growth factor beta superfamily and has a striking sequence similarity to the decapentaplegic locus in Drosophila melanogaster, a major determinant of pattern specification during embryogenesis. BMP2A is thought to be involved in cartilage and bone formation during embryogenesis, but may have additional functions in morphogenesis as implied by its expression in various organs and embryonic tissues of mice. Human BMP2A, assigned to chromosome 20 by the use of human-Chinese hamster ovary cell hybrids, is considered to be a reasonable candidate gene for the autosomal dominant disease of fibrodysplasia (myositis) ossificans progressiva. We have confirmed the localization of BMP2A to chromosome 20 and regionally assigned the locus to 20p12 by radioactive and nonradioactive in situ hybridization.

The polymorphic DNA marker D20S14 was previously mapped to human chromosome 20 and shown to be linked to two other DNA markers, D20S5 and D20S6, located at 20p12. This segment has been implicated in several human diseases. Because of its importance, we mapped the D20S14 locus to 20p12----p11.2 by radioactive in situ hybridization.

Two posterior pituitary hormones oxytocin and arginine-vasopressin control the important activities of water excretion, parturition and lactation. Both these hormones are synthesized as inactive precursors in the hypothalamus along with their carrier proteins neurophysin I and neurophysin II respectively and are activated upon transport to posterior pituitary. Human genes for both oxytocin-neurophysin I (OXT) and arginine-vasopressin-neurophysin II (ARVP) are cloned and found to be linked on chromosome 20 separated by approximately 12 kb of intergenic sequences. Though OXT is not yet associated with any disease, ARVP is linked to the autosomal dominant disease neurohypophyseal diabetes insipidus (AD-NDI). We have mapped regionally the OXT locus to chromosome 20p13 by both radioactive (ISH) and fluorescence in situ hybridization (FISH).

Endothelin 3 is a novel vasoactive peptide of unique structure and belongs to the endothelin gene family. Though it mainly functions as a vasoconstrictor/pressor, it plays a major role in a wide variety of other biological functions, possibly as a novel neuropeptide. Studies with human-mouse somatic cell hybrids have suggested that the gene EDN3 is localized on human chromosome 20. We confirmed this localization and regionally mapped the gene to the region 20q13.2-q13.3.

Ribophorin I and II (RPN I and RPN II), two specific glycoproteins, span the rough regions of the endoplasmic reticulum (RER) and are thought to play an important role either in translocation or in the maintenance of RER. Studies with human-mouse somatic cell hybrids have localized the gene for RPN I on human chromosome 3q, while RPN II is on chromosome 20. Using a radioactive labelled cDNA probe, we have regionally mapped the RPN II gene to human chromosome 20q12-q13.1 by in situ hybridization. This assignment predicts a location of the murine homologue, Rpn-2, to the syntenic segment on mouse chromosome 2 in close proximity to Ada, Src and Gnas.

Two forms of urinary trypsin inhibitor, A and B, were purified from the urine of pregnant women. Form A was the only inhibitor present in fresh urine and inhibitor B arose from degradation of A upon storage of urine. The molecular masses of A and B were about 44 and 20 kDa, respectively, as judged from dodecyl-sulfate polyacrylamide gel electrophoresis, but about 60 kDa and 30 kDa, respectively, as judged from gel filtration analysis. The discrepancy can perhaps be explained by the carbohydrate content amounting to about 10% of each inhibitor. After reduction with mercaptoethanol, inhibitor A and inhibitor B had identical apparent molecular masses of about 20 kDa on dodecyl-sulfate gel electrophoresis. These results and the results of amino acid analysis suggest that one molecule of inhibitor A yields two molecules of inhibitor B. On agarose gel electrophoresis inhibitor A migrated as a rather broad band in the prealbumin region and inhibitor B as 3 well defined bands in the beta-region. Specific antisera were raised against inhibitor A and B. The two inhibitors showed the immunologic reaction of identity with each other and with the plasma inter-alpha-trypsin inhibitor, when using either antiserum. The inhibitors both gave quantitative inhibition of bovine trypsin, the results indicating a 4/1 trypsin/inhibitor molar ratio for A and a 2/1 ratio for B. The two substances also effectively inhibited granulocyte elastase. No inhibition of porcine pancreatic elastase was demonstrable.