Biomedical subjects

U Pohl

Publications and source records attributed to U Pohl.

At least 19 recordsLinked to original sources

PubMed2001

Endothelium-specific replacement of the connexin43 coding region by a lacZ reporter gene.

The murine gap junction protein connexin43 (Cx43) is expressed in blood vessels, with vastly different contribution by endothelial and smooth muscle cells. We have used the Cre recombinase under control of TIE2 transcriptional elements to inactivate a floxed Cx43 gene specifically in endothelial cells. Cre-mediated deletion led to replacement of the Cx43 coding region by a lacZ reporter gene. This allowed us to monitor the extent of deletion and to visualize the endothelial expression pattern of Cx43. We found widespread endothelial expression of the Cx43 gene during embryonic development, which became restricted largely to capillaries and small vessels in all adult organs examined. Mice lacking Cx43 in endothelium did not exhibit altered blood pressure, in contrast to mice deficient in Cx40. Our results show that lacZ activation after deletion of the target gene allows us to determine the extent of cell type-specific deletion after phenotypical investigation of the same animal.

Animals↗

PubMed2001

Hypoxia-induced upregulation of eNOS gene expression is redox-sensitive: a comparison between hypoxia and inhibitors of cell metabolism.

Several papers report a hypoxia-induced upregulation of the endothelial nitric oxide synthase (eNOS) mRNA expression. Since there is no known hypoxia-sensitive element binding site in the eNOS promoter, we reasoned that the effect of hypoxia could be simulated by a metabolically elicited alteration of the redox state. Therefore, cultured porcine aortic endothelial cells (PAEC) were exposed to hypoxia (1-10% O(2)) or inhibitors of cellular energy metabolism including rotenone, 2, 4 dinitrophenol (DNP) and 2-deoxyglucose for 6 to 24 h. Additionally, cells were treated with lactate and nicotinic acid to alter the cellular NAD(P)H/NAD(P) ratio without changes of energy supply. The cellular NAD(P)H/NAD(P) ratio was used as an index of the cellular redox state and determined using the MTT-assay. Hypoxia increased eNOS mRNA transcription and MTT-reduction in a manner inversely proportional to pO(2). Exposure to rotenone, DNP, and lactate increased the NAD(P)H/NAD(P) ratio, MTT-reduction, and eNOS mRNA also in parallel. In contrast, 2-deoxyglucose and nicotinic acid attenuated both MTT-reduction and eNOS mRNA expression. In order to study a potential role of the redox regulated transcription factor complex AP-1 in hypoxia-induced eNOS mRNA transcription, c-jun expression was determined and decoy experiments were performed. c-jun expression paralleled changes of eNOS mRNA expression and MTT-reduction. Furthermore, in the presence of oligodeoxynucleotides corresponding to the AP-1 binding sites of the eNOS promoter, the hypoxia and chemically induced eNOS mRNA expression was completely abolished. We propose that hypoxia, by altering cellular metabolism, leads to an increase in the cellular NAD(P)H/NAD(P) ratio which favors enhanced eNOS expression by redox-sensitive AP-1 mediated transcriptional control.

Animals↗

PubMed2001

[Examination of body concept of female patients with borderline personality disorder].

The operationalized diagnosis of borderline personality disorder (BPD) by DSM-IV and ICD-10 (research criteria) is meanwhile considered as valid and reliable. A disturbed or inappropriate body image within the group of borderline patients is not assessed by these instruments. Using the "Frankfurter Körperkonzeptskalen" [6], we compared the body concept of 47 female patients with BPD with a group of patients with the diagnosis of Bulimia nervosa and with a representative sample of healthy subjects. We found highly significant differences between patients with BPD and the clinical controls as well as to the norm group. Neither dissociative symptoms nor sexual child abuse correlate with the extent of the negative body concept. Correlations between self-concept (Giessen-Test) and body concept as well as clinical implications of these findings are discussed.

Adult↗

PubMed2001

A randomized evaluation of a computer-based nursing documentation system.

A two-month randomized, controlled trial based on 60 patients has been performed on a ward of the Department of Psychiatry at Heidelberg University Medical Center, Germany, to investigate the influence of computer-based nursing documentation on time investment for documentation, quality of documentation and user acceptance. Time measurements, questionnaires, documentation analysis and interviews were used to compare patients documented with the computer-based system (PIK group) with the control group (patients documented with the paper-based system). The results showed the advantages and disadvantages of computer-based nursing documentation. Time needed for nursing care planning was lower in the PIK group. Some formal aspects of quality were considerably better in the PIK group. On the other hand, time required for documentation of tasks and for report writing was greater in the PIK group. User acceptance increased significantly during the study. The interviews indicated a positive influence of PIK on the cooperation between nurses and physicians.

Attitude to Computers↗

PubMed2001

Analysis and modelling of the multi-professional treatment process: preliminary results.

This paper presents first results of a research project aimed at improving co-operative work initiatives in hospitals. A holistic analysis of the treatment process is presented as a precondition for process reengineering, quality measurements and improvement of multi-professional co-operation. Treatment process modelling attempts within the last years have concentrated on specialised points of views, such as business process modelling or communication modelling. In contrast, we have developed a framework consisting of several views of the treatment process. We tested our framework in a broad system analysis within the Department of Child and Adolescent Psychiatry of the Heidelberg University Hospitals. Our preliminary results support the framework. Weaknesses were described precisely in both the field of organisational procedures and information management.

Comprehensive Health Care↗

PubMed2000

Oxidized LDL increases the sensitivity of the contractile apparatus in isolated resistance arteries for Ca(2+) via a rho- and rho kinase-dependent mechanism.

BACKGROUND: Oxidized LDL reduces NO-mediated and endothelium-derived hyperpolarizing factor-mediated dilations. We studied, in hamster skeletal muscle resistance arteries (213+/-8 micrometer n=51), whether an altered vascular smooth muscle (VSM) response, particularly sensitization of the VSM contractile apparatus to Ca(2+), is involved in this oxLDL effect. Methods and Results-VSM or endothelial [Ca(2+)](i) and vascular diameter were measured in response to norepinephrine (0.3 micromol/L), sodium nitroprusside (10 micromol/L), C-type natriuretic peptide (1 to 100 nmol/L), papaverine (0.1 to 10 micromol/L), or the endothelial agonist acetylcholine (ACh, 0.01 to 1 micromol/L). OxLDL significantly increased resting VSM [Ca(2+)](i) (11+/-3%), decreased diameter (8+/-2%), and enhanced norepinephrine-induced constrictions. Dilations to sodium nitroprusside and C-type natriuretic peptide were significantly reduced (by 10+/-2% and 35+/-6%), whereas dose-response curves for papaverine and ACh were shifted to the right, despite unchanged increases in endothelial Ca(2+) after ACh. OxLDL significantly shifted the Ca(2+)-diameter relation to the left, as assessed by stepwise increasing extracellular Ca(2+) (0 to 3 mmol/L) in depolarized skeletal muscle resistance arteries. This sensitization to Ca(2+) by oxLDL was abolished after inhibition of Rho (C3 transferase) or Rho kinase (Y27632). CONCLUSIONS: OxLDL reduces VSM responsiveness to vasodilators by increasing VSM Ca(2+) but preferentially by sensitizing VSM to Ca(2+) via a Rho- and Rho kinase-dependent pathway.

Acetylcholine↗

PubMed2000

The small G-protein Rac mediates depolarization-induced superoxide formation in human endothelial cells.

Superoxide anions impair nitric oxide-mediated responses and are involved in the development of hypertensive vascular hypertrophy. The regulation of their production in the vascular system is, however, poorly understood. We investigated whether changes in membrane potential that occur in hypertensive vessels modulate endothelial superoxide production. In cultured human umbilical vein endothelial cells, changes in membrane potential were induced by high potassium buffer, the non-selective potassium channel blocker tetrabutylammonium chloride (1 mm), and the non-selective cation ionophore gramicidin (1 micrometer). Superoxide formation was significantly elevated to a similar degree by all three treatments (by approximately 60%, n = 23, p < 0.01), whereas hyperpolarization by the K(ATP) channel activator Hoe234 (1 micrometer) significantly decreased superoxide formation. Depolarization also induced an increased tyrosine phosphorylation of several not yet identified proteins (90-110 kDa) and resulted in a significant increase in membrane association of the small G-protein Rac. Accordingly, the Rac inhibitor Clostridium difficile toxin B blocked the effects of depolarization on superoxide formation. The tyrosine kinase inhibitor genistein (30 micrometer, n = 15) abolished depolarization-induced superoxide formation and also prevented depolarization-induced Rac translocation associated with it. It is concluded that depolarization is an important stimulus of endothelial superoxide production, which involves a tyrosine phosphorylation-dependent translocation of the small G-protein Rac.

Endothelium, Vascular↗

PubMed2000

Impaired conduction of vasodilation along arterioles in connexin40-deficient mice.

Connexins have been hypothesized to play an important role in intercellular communication within the vascular wall and may provide a mechanistic explanation for conduction of vasomotor responses. To test this hypothesis, we studied the transmission of vasomotor responses in the intact skeletal muscle microcirculation of connexin40-deficient mice (Cx40(-/-)). Arterioles were locally stimulated with hyperpolarizing dilators (acetylcholine [ACh] as well as bradykinin [Bk]) or depolarizing K(+) solution, and the resulting changes in diameter were measured using a videomicroscopy technique at the site of application and up to 1.32 mm upstream. Arterial pressure was elevated 25% in Cx40(-/-) mice (94+/-5 versus 75+/-4 mm Hg). Vessels selected for study had equivalent basal diameter and vasomotor tone in both genotypes of mice. Vasomotion was present in small arterioles of both genotypes, but its intensity was exaggerated in Cx40(-/-) mice. ACh and Bk induced dilation (33% and 53%, respectively, of maximal response) at the site of application that was of similar magnitude in both genotypes. These dilations were observed to spread upstream within <1 second without significant attenuation in Cx40(+/+) mice. However, spreading was severely attenuated in Cx40(-/-) animals (11+/-4% versus 35+/-7% with ACh and 38+/-5% versus 60+/-7% with Bk in Cx40(-/-) and Cx40(+/+), respectively; P<0.05). In contrast, conducted vasoconstrictions, induced by K(+) solution decreased equally with distance in both genotypes. These results support a significant role for Cx40 in vascular intercellular communication. Our observations indicate that Cx40 is required for normal transmission of endothelium-dependent vasodilator responses and may underlie altered vasomotion patterns.

Acetylcholine↗

PubMed2000

A transcript map of the chromosome 19q-arm glioma tumor suppressor region.

Allelic loss of the chromosome 19q arm is a frequent event in human diffuse gliomas, suggesting that it contains a tumor suppressor gene. Recent deletion mapping studies have broadly implicated a 1.6-Mb interval between D19S241E and D19S596, with a limited subset of tumors, suggesting that the region may be as narrow as 150 kb. Focusing on this smaller interval, we have used cDNA selection, exon amplification, and genomic sequencing to identify three novel transcripts (EHD2, GLTSCR1, and GLTSCR2) and to map two known genes (SEPW1 and CRX). A partial transcript map of 19 transcripts and two EST markers has been constructed for the 1.6-Mb interval D19S241E-D19S596. Ten of these transcripts, including the 5 mapped to the 150-kb deletion interval, have been examined for alterations in a panel of gliomas with allelic loss of 19q. Tumor-specific alterations have not been identified in the transcripts examined thus far. Collectively, these data should facilitate subsequent efforts to identify and characterize the remaining transcripts in the 1.6-Mb interval.

Base Sequence↗

PubMed2000

EHD2, EHD3, and EHD4 encode novel members of a highly conserved family of EH domain-containing proteins.

Exon trapping from a bacterial artificial chromosome (BAC 78138) mapping to the 19q13.3 glioma tumor suppressor candidate region yielded two exons that recognized a 3.6-kb transcript on Northern blot. Screening of a human fetal brain cDNA library with these exons identified three novel genes, designated EHD2, EHD3, and EHD4, which are homologous to the recently characterized human EHD1 (testilin/HPAST) and its mouse homolog Ehd1, as well as to homologs in Drosophila (Past1) and Caenorhabditis elegans. Alignment of the predicted peptide sequences revealed striking similarities, with multiple conserved regions that include a nucleotide-binding consensus site at the N-terminus, a bipartite nuclear localization signal, and an eps15 homology (EH) protein-binding domain with an EF-hand motif at the C-terminus. The genes are specifically expressed, with EHD2 highly expressed in heart, EHD3 in brain and heart, and EHD4 in heart and pancreas. EHD2 was confirmed to originate from BAC 78138 at 19q13.3; radiation hybrid mapping localized EHD3 and EHD4 to 2p21 and 15q11.1, respectively; EHD1 has been previously mapped to 11q13. The three EHD1 paralogs therefore represent novel members of a family of human EH domain-containing proteins that may play a role in endocytosis and signaling. Mutation analysis of the five coding exons of EHD2 in gliomas failed to detect any tumor-specific alterations, thus indicating that EHD2 is an unlikely candidate for the 19q tumor suppressor gene.

Amino Acid Sequence↗

PubMed2000

Mapping of the chromosome 19 q-arm glioma tumor suppressor gene using fluorescence in situ hybridization and novel microsatellite markers.

Allelic loss of chromosome arm 19q is a frequent event in human diffuse glioma, suggesting the presence of a tumor suppressor gene. Previous loss of heterozygosity (LOH) analyses have mapped this gene to a 1.4-megabase interval, between the genetic markers D19S412 and STD. Further narrowing of this interval has been limited by the resolution of mapped polymorphic markers. In the present study, we have used genomic clones mapped to 19q as fluorescence in situ hybridization (FISH) probes to map the breakpoints of 13 gliomas with 19q13.3 deletion boundaries. In addition, we have developed three new polymorphic microsatellite markers (D19S1180, D19S1181, and D19S1182) that map between D19S412 and STD and have used these new markers to identify two gliomas with small deletions between the D19S412 and STD markers. Collectively, these data suggest that the region of common deletion may be as narrow as 150 kb and should facilitate future efforts to identify the glioma 19q tumor suppressor gene.

Chromosome Deletion↗

PubMed2000

Interaction of endothelial autacoids in microvascular control.

The endothelium releases dilating autacoids, such as nitric oxide (NO), prostaglandins, and endothelium-derived hyperpolarizing factor (EDHF). Different mechanisms of interaction between these autacoids have been identified and are briefly reviewed here. NO, which dilates resistance vessels via the second messenger cGMP, also amplifies the responses to vasodilators, which act by elevation of cAMP (e.g., prostaglandins), in a synergistic manner. This amplification is most likely due to a cGMP-dependent inhibition of the breakdown of cAMP. An interaction of these autacoids can not only be found in vascular smooth muscle cells but also in the endothelium. A closer look towards the site of production reveals that prostacyclin attenuates the release of NO, which is achieved by a decrease of the intracellular calcium levels in endothelial cells. This feedback mechanism leads to enhanced NO formation after inhibition of cyclooxygenase. While NO does not seem to modulate prostaglandin synthesis, it has been shown recently in coronary arteries that NO attenuates the release of EDHF. Under conditions of compromised NO formation EDHF might act via this mechanism as a second line of defense. This could help in maintaining endothelium-dependent vasodilation unless an altered smooth muscle responsiveness generally reduces the efficacy of endothelial vasodilators (including EDHF). Convincing experimental evidence for such an impaired relaxation of smooth muscle has been presented recently following exposure to elevated oxLDL levels.

Animals↗

PubMed2000

Evaluation of inpatient dialectical-behavioral therapy for borderline personality disorder--a prospective study.

Dialectical-Behavioral Therapy for Borderline Personality Disorder (DBT) developed by M. Linehan is specifically designed for the outpatient treatment of chronically suicidal patients with borderline personality disorder. Research on DBT therapy, its course and its results has focused to date on treatments in an outpatient setting. Hypothesizing that the course of therapy could be accelerated and improved by an inpatient setting at the beginning of outpatient DBT, we developed a treatment program of inpatient therapy for this patient group according to the guidelines of DBT. It consists of a three-month inpatient treatment prior to long-term outpatient therapy. In this pilot study 24 female patients were compared at admission to the hospital, and at one month after discharge with respect to psychopathology and frequency of self-injuries. Significant improvements in ratings of depression, dissociation, anxiety and global stress were found. A highly significant decrease in the number of parasuicidal acts was also reported. Analysis of the average effect sizes shows a strong effect which prompts the development of a randomized controlled design.

Adult↗

PubMed2000

Differential role of angiotensin II receptor subtypes on endothelial superoxide formation.

The physiological role of the angiotensin II AT2 receptor subtype is not fully characterized. We studied whether AT2 receptor could antagonize AT1 mediated superoxide formation in endothelial cells. In quiescent human umbilical vein endothelial cells (HUVEC) superoxide formation was measured after long-term incubation (6 h) with angiotensin II in the presence or absence of its receptor blocker candesartan (AT1) or PD123319 (AT2) using the cytochrome c assay. In separate experiments, the effects of AT2 mediated effects on activities of cellular phosphates including the src homology 2 domain containing phosphatases (SHP-1) was studied. The basal superoxide formation (0.19+/-0.03 nmol superoxide mg protein(-1) min(-1)) in HUVEC was increased by 37.1% after exposure to angiotensin II (100 nM,) which was due to an activation of a NAD(P)H oxidase. This was abolished by candesartan (1 microM) as well as the tyrosine kinase inhibitor genistein. In contrast, blockade of AT2 receptors by PD123319 enhanced the superoxide formation by 73.7% in intact cells. Stimulation of AT2 went along with an increased activity of tyrosine phosphatases in total cell lysates (29.8%) and, in particular, a marked stimulation of src homology 2 domain containing phosphatases (SHP-1, by 293.4%). The tyrosine phosphatase inhibitor vanadate, in turn, prevented the AT2 mediated effects on superoxide formation. The expression of both angiotensin II receptor subtypes AT1 and AT2 was confirmed by RT - PCR analysis. It is concluded that AT2 functionally antagonizes the AT1 induced endothelial superoxide formation by a pathway involving tyrosine phosphatases.

Cells, Cultured↗

PubMed2000

Chronic increases in transmural pressure reduce NO-mediated dilations in isolated resistance arteries of the hamster.

It is unclear whether the impairment of NO-mediated dilation in hypertension is the cause or the consequence of high blood pressure. We therefore studied in isolated resistance arteries whether elevated transmural pressure affects NO-mediated dilation. Arteries (n=5-7) were perfused at hydrostatic pressures of either 45, 120 or 160 mmHg for 48 h. Subsequently, diameter and calcium responses (fura 2) were studied at a transmural pressure of 45 mmHg. Pre-exposure to 120 and 160 mmHg reduced resting diameters and minimal diameters after stimulation with noradrenaline and significantly increased corresponding intracellular free calcium levels in vascular smooth muscle. Moreover, the NO-mediated dilation in response to acetylcholine was significantly reduced although the increase in endothelial calcium was not altered. Dilations induced by the NO donor SNP were not affected. It is concluded that chronically elevated pressure per se impairs endothelial NO production by a mechanism distal to receptor-dependent calcium increases.

Animals↗

PubMed2000

Large arterioles in the control of blood flow: role of endothelium-dependent dilation.

Although it is generally assumed that small arterioles form the major site of vascular resistance, microcirculatory studies revealed that 40-55% of the total network resistance can reside in large arterioles and small arteries. Thus, the mechanisms that control smooth muscle tone in these vessels have a major impact on the overall conductance of the vascular network. These control mechanisms are different from those in small arterioles: Aside from an apparently reduced sensitivity to metabolites, the large resistance vessels are normally too far away from the capillary areas which they feed to be reached by diffusing metabolites from dependent cells within a reasonable period of time. Rather, recent intravital microscopic studies suggest that large resistance vessels are under tight control of endothelial factors such as nitric oxide and endothelium-derived hyperpolarising factor (EDHF). Nitric oxide opposes myogenic constrictions of large arterioles that potentially would impair tissue perfusion and oxygenation. Moreover, nitric oxide and EDHF play an important role in the co-ordination of large and small resistance vessel behaviour that is pivotal for the adaptation of blood flow to altered tissue oxygen demands.

Animals↗

PubMed2000

[Recent developments in systematic evaluation of the therapeutic process. Scales for the assessment of adherence and competence].

The assessment of therapists' behaviour an essential part of psychotherapy research, especially for controlling the validity of studies. In the last decade, particularly in English-speaking countries, research has been developed that studied two constructs important in this context: adherence and competence. Adherence refers to the extent to which therapists use the strategies recommended in a therapy manual; competence refers to the quality of the use of these strategies. This article is a review of research associated with adherence and competence scales and its relevance. First of all, definitions and examples of use are presented and discussed. Secondly, some of the most important scales that have been developed in this context are described. Thirdly, aspects of realisation are presented. Finally, the pros and cons of this type of research are discussed. It is argued that despite some problems adherence and competence scales are a significant contribution to the systematic assessment of therapist behaviour.

Germany↗

PubMed2000

Pitfalls of using lucigenin in endothelial cells: implications for NAD(P)H dependent superoxide formation.

Since an increased endothelial superoxide formation plays an important role in the pathogenesis of endothelial dysfunction its specific detection is of particular interest. The widely used superoxide probe lucigenin, however, has been reported to induce superoxide under certain conditions, especially in the presence of NADH. This raises questions as to the conclusion of a NAD(P)H oxidase as the major source of endothelial superoxide. Using independent methods, we showed that lucigenin in the presence of NADH leads to the production of substantial amount of superoxide (approximately 15-fold of control) in endothelial cell homogenates. On the other hand, these independent methods revealed that endothelial cells without lucigenin still produce superoxide in a NAD(P)H-dependent manner. This was blocked by inhibitors of the neutrophil NADPH oxidase diphenyleniodonium and phenylarsine oxide. Our results demonstrate that a NAD(P)H-dependent oxidase is an important source for endothelial superoxide but the latter, however, cannot be measured reliably by lucigenin.

Acridines↗