Biomedical subjects
M Mueller
Publications and source records attributed to M Mueller.
Solution structure of a POU-specific homeodomain: 3D-NMR studies of human B-cell transcription factor Oct-2.
The POU DNA-binding motif defines a conserved family of eukaryotic transcription factors involved in regulation of gene expression. This bipartite motif consists of an N-terminal POU-specific domain (POUs), a flexible linker, and a C-terminal POU-specific homeodomain (POUHD). Here we describe the solution structure of a POU-specific homeodomain. An NMR model is obtained from Oct-2, a human B-cell specific transcription factor which participates in the regulation of immunoglobulin genes. A fragment of Oct-2 containing POUHD and an adjoining linker was expressed in Escherichia coli and characterized by three-dimensional nuclear magnetic resonance (3D-NMR) spectroscopy. Complete 1H and 15N resonance assignment of the POUHD moiety is presented. The POUHD solution structure, as calculated by distance geometry and simulated annealing (DG/SA), is similar to that of canonical homeodomains. A salient difference between solution and crystal structures is observed in the C-terminal segment of alpha-helix 3 (the HTH recognition helix), which is not well ordered in solution. Because this segment presumably folds upon specific DNA binding, its flexibility in solution may reduce the intrinsic DNA affinity of POUHD in the absence of POUs.
CGP 48664, a new S-adenosylmethionine decarboxylase inhibitor with broad spectrum antiproliferative and antitumor activity.
Inhibitors of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (SAMDC), derived from methylglyoxal-bis(guanylhydrazone) (MGBG), have been shown to have significant antitumor activity in several human solid tumor systems (U. Regenass et al., Cancer Res., 52:4712-4718, 1992). From an ongoing effort to synthesize derivatives with increased enzyme specificity and potency and improved antitumor efficacy, we have now identified CGP 48664, a 4-amidinoindan-1-one 2'-amidinohydrazone (J. Stanek et al., J. Med. Chem., 36:2168-2171, 1993). The compound displays potent inhibition of SAMDC (50% inhibitory concentration, 5 nM), modest inhibition of diamine oxidase (50% inhibitory concentration, 4 microM), and no detectable inhibition of ornithine decarboxylase. CGP 48664 inhibits the growth of a panel of human and mouse tumor cell lines, including one which expresses the multidrug resistance phenotype, with 50% inhibitory concentrations ranging between 0.3 and 3 microM. CGP 48664 does not seem to utilize the polyamine transport carrier system since it competes poorly with spermidine for uptake into L1210 cells (Ki 161 microM) and inhibits the growth of polyamine transport-deficient Chinese hamster ovary cells. Relative to MGBG or previously described MGBG analogues, CGP 48664 accumulates to much lower intracellular concentrations. Treatment of the L1210 cell for 48 h with 3 microM CGP 48664 decreases SAMDC activity to < 10% of control and initiates a compensatory 3-fold rise in ornithine decarboxylase. Consistent with SAMDC inhibition, putrescine pools increase 10-fold, whereas spermidine and spermine pools fall to < 10% of control. In contrast to MGBG, CGP 48664 displays attenuated antimitochondrial activity as indicated by a lack of effect on pyruvate oxidation and mitochondrial DNA levels under treatment conditions which inhibit cell proliferation. Specificity of drug action was indicated further by prevention of L1210 cell growth inhibition by exogenous spermidine or spermine. More convincingly, Chinese hamster ovary cells made approximately 1000-fold resistant by chronic exposure to the analogue were found to selectively overexpress SAMDC mRNA due to gene amplification. The new SAMDC inhibitor showed potent antitumor activity against syngeneic tumors (B16 melanoma and Lewis lung carcinoma) and nude mouse human tumor xenografts (T-24 bladder carcinoma, SK MEL-24 melanoma, and MALME-3M melanoma). On the basis of its novel structure, its apparent specificity of action, and its potent antitumor activity, CGP 48664 is the candidate drug for further preclinical development.
Changes in arterial wall compliance after endovascular stenting.
PURPOSE: The response of arterial wall to endovascular stenting after angioplasty is not well understood. Additionally, changes in the elastic properties of stented vessels are unknown in situ. Vascular compliance was measured in normal canine iliac arteries (n = 11) before and after intravascular ultrasound-guided deployment of self-expandable metallic stents. METHODS: Nine animals were restudied and killed 1, 2, and 4 weeks after initial deployment, and two dogs were studied at deployment only. An absolute induction angiometer was used to make in situ measurements of vessel compliance via catheter-based delivery. The angiometer consists of a wire loop probe, which conforms to the diameter of the vessel in which it is placed. Systolic/diastolic changes in loop diameter are translated into measureable changes of induced voltage. RESULTS: Mean compliance of the artery before and immediately after stenting was 4.4 +/- 2.1 and 1.9 +/- 2.0 (x 10(-2) diameter %/mm Hg), respectively. As early as 1 week after deployment, stented arteries began to lose expansile properties, and some were noncompliant. At explantation diminished compliance was accompanied by a periadventitial fibrous reaction around stented vessels. A thin, unobstructing layer of neointimal hyperplasia covered the iliac stents at all intervals, and all vessels remained patent and free of thrombus. CONCLUSIONS: The potential advantages provided by a flexible, radially compliant stent are lost within a relatively short time after implantation in nonatherosclerotic canine arteries.
Traumatic coronary sinocameral fistula from a penetrating cardiac injury: case report and review of the literature.
Clinically apparent penetrating cardiac wounds require emergent surgical therapy without the theoretical benefit of preoperative evaluation apart from the physical examination. Residual intracardiac injuries discovered following the successful resuscitation and repair of immediate life-threatening cardiac wounds should be sought in survivors of chest trauma who demonstrate new regurgitant murmurs or evidence of congestive heart failure. To our knowledge, we report here the second case of a traumatic left ventricle-to-coronary sinus fistula, along with recommendations for its management.
Effect of filler content of restorative resins on retentive strength to acid-conditioned enamel.
In this in vitro study the retentive strength of enamel, either conventionally etched with a phosphoric acid gel or conditioned with an oxalic acid containing etching agent, was investigated. Five experimental restorative resins with filler contents between 0 and 78% by weight were polymerized on the etched enamel surfaces. In spite of the pronounced micromorphological differences of the patterns generated with the two etching agents, the bond strengths mediated were very similar. Application of an intermediate low-viscous or adhesive resin had no influence on bond strength. Exponential relationships were found between the filler content of the resins on the one hand and their flexural moduli and flexural strengths on the other. Significant exponential relations were also demonstrated between filler content and bond strength to the etched enamel surface. The linear relationship between mechanical properties of the resins and their bond strengths to etched enamel indicates that debonding occurred by cohesive failure in resin.
[Does laparoscopic lysis of adhesions make sense?].
Between May 1989 and December 1991 23 patients were operated on laparoscopically for symptomatic peritoneal adhesions. 19 were followed up by telephone using a standardized questionnaire an average of 18.3 months post-operatively (range 5-36 months). 12 patients were totally painfree, 3 complained of slight pain, 2 patients of fairly severe and 2 of severe pain. 15 patients considered the outcome of the operation to be good or fairly good and 18 said they would undergo the same operation in similar circumstances. We therefore recommend laparoscopic adhesiolysis in cases of acute or chronic abdominal pain, provided other causes of abdominal discomfort have been ruled out.
Mass spectrometric identification and quantitation of arachidonate-containing phospholipids in pancreatic islets: prominence of plasmenylethanolamine molecular species.
D-Glucose induces insulin secretion from beta-cells of pancreatic islets by processes involving glycolytic metabolism and generation of ATP. Glucose also induces hydrolysis of beta-cell membrane phospholipids and accumulation of nonesterified arachidonate, which facilitates Ca2+ entry and the rise in beta-cell Ca2+ concentration that is a critical signal in the induction of insulin secretion. Glucose-induced hydrolysis of arachidonate from beta-cell phospholipids is mediated in part by an ATP-stimulated, Ca(2+)-independent (ASCI)-phospholipase A2 (PLA2), which, in vitro, prefers plasmalogen over diacylphospholipid substrates, but it is not known whether islets contain plasmalogens. We have identified and quantitated the major species of arachidonate-containing phospholipids in pancreatic islets by high-performance liquid chromatographic and mass spectrometric analyses. Arachidonate has been found to constitute 30% of the total islet glycerolipid fatty acyl mass. Ethanolamine phospholipids contain 30% of total islet arachidonate, and 44% of that amount resides in three plasmenylethanolamine molecular species with residues of palmitic, oleic, or stearic aldehydes in the sn-1 position. These endogenous islet plasmenylethanolamine species are hydrolyzed more rapidly than phosphatidylethanolamine species by islet ASCI-PLA2 in vitro and are also hydrolyzed in intact islets stimulated with secretagogues. ASCI-PLA2-catalyzed hydrolysis of islet plasmenylethanolamine species in vitro is inhibited by a selective haloenol lactone suicide substrate (HELSS) which is sterically similar to plasmalogens, and HELSS also inhibits all temporal phases of both eicosanoid release and insulin secretion from secretagogue-stimulated pancreatic islets. Islet beta-cell ASCI-PLA2-catalyzed hydrolysis of arachidonate from endogenous plasmenylethanolamine substrates may be an intermediary biochemical event in the induction of insulin secretion.
Portal magnetic resonance angiography.
MR angiography has shown definite clinical use in the portal venous system. Methods have been developed for noninvasive assessment of portal venous anatomy and blood flow using a variety of techniques. Time-of-flight techniques for portal angiography and both time-of-flight and phase-contrast techniques for flow measurement are reviewed.
Influence of aluminum oxalate solutions acidity and conditioning times on resin bond strength to enamel.
The present investigation demonstrated that variations in pH between 0.8 and 2.1 and in the duration of etching with an aluminum oxalate/glycine conditioning solution between 5 and 120 seconds has no influence on the retentive strength of restorative polymer to enamel, although the micromorphological appearance of enamel is clearly affected. The lower the pH of and the longer the etching time with the conditioning solution, the more pronounced is the roughness pattern generated. SEM inspection of the fracture sites following debonding of shear bond strength specimens verified consistently cohesive failure in resin close to the interface.
Endoscopic laser therapy in a case of gastric antral vascular ectasia (watermelon stomach).
We report the case history of a 75-year-old female patient suffering from extensive gastric antral vascular ectasia (watermelon stomach) with portal hypertension in alcoholic liver cirrhosis. Iron deficiency anaemia, due to chronic blood loss from the antral lesions, required repeated transfusions. Conservative treatment failed, but a surgical intervention (antrectomy) was discarded in view of concomitant diseases. Endoscopic laser photocoagulation therapy with a Nd:YAG-laser eradicated the characteristic antral lesions, which was accompanied by a significant reduction of transfusion requirements. No relapse has been observed since then. Endoscopic laser photocoagulation appears to be an efficacious and safe alternative in the treatment of gastric antral vascular ectasia.
Selective inhibition of tumor cell growth by a recombinant single-chain antibody-toxin specific for the erbB-2 receptor.
A high percentage of human breast and ovarian tumors display amplified c-erbB-2 gene copies, leading to overexpression of the growth factor receptor. Its membrane location and elevated expression make the erbB-2 protein an appropriate target for a directed tumor therapy. We have used recombinant DNA technology to produce a single-chain antibody-exotoxin A (scFv-ETA) fusion protein which specifically binds the human erbB-2 receptor. The scFv portion is composed of the heavy- and light-chain variable domains of a monoclonal antibody which recognizes the extracellular domain of the human erbB-2 receptor. The bacterially produced scFv-ETA protein was shown to bind specifically to cells expressing the human erbB-2 protein. The scFv-ETA inhibits protein synthesis in erbB-2-expressing tumor cells at doses ranging from 2 to 200 ng/ml and is cytotoxic for these cells at equivalent doses. In athymic nude mice, administration of the scFv-ETA inhibited the growth of erbB-2-overexpressing human ovarian carcinoma cells.
Arachidonic acid metabolism in isolated pancreatic islets. VI. Carbohydrate insulin secretagogues must be metabolized to induce eicosanoid release.
Pancreatic islets stimulated with D-glucose are known to liberate arachidonic acid from membrane phospholipids and release prostaglandin E2 (PGE2). A component of the eicosanoid release induced by D-glucose has been demonstrated to occur without calcium influx and must be triggered by other coupling mechanisms. In this study, we have attempted to identify mechanisms other than calcium influx which might couple D-glucose stimulation to hydrolysis of arachidonate from membrane phospholipids in islet cells. We have found that occupancy of the beta cell plasma membrane D-glucose transporter is insufficient and that D-glucose metabolism is required to induce islet PGE2 release because 3-O-methylglucose fails to induce and mannoheptulose prevents PGE2 release otherwise induced by 17 mM D-glucose. The carbohydrate insulin secretagogues mannose and D-glyceraldehyde have also been found to induce islet PGE2 release, but the non-secretagogue carbohydrates L-glucose and lactate do not. Carbohydrate secretagogues are known to be metabolized to yield ATP and induce depolarization of the beta cell plasma membrane. We have found that depolarization by 40 mM KCl induces PGE2 release only in the presence and not in the absence of extracellular calcium, but exogenous ATP induces islet PGE2 release with or without extracellular calcium. Carbachol is demonstrated here to interact synergistically with increasing concentrations of glucose to amplify PGE2 release and insulin secretion. Pertussis toxin treatment is shown here not to prevent PGE2 release induced by glucose or carbachol but to increase the basal rate of PGE2 release and the islet cyclic AMP content. Theophylline (10 mM) exerts similar effects. Eicosanoid release in pancreatic islets can thus be activated by multiple pathways including muscarinic receptor occupancy, calcium influx, increasing cAMP content, and a metabolic signal derived from nutrient secretagogues, such as ATP.
Does immobilization influence the systemic acceleratory phenomenon that accompanies local bone repair?
The rate of remodeling in the region of a bone defect exceeds normal tissue activity. It was Frost who described this reaction as the regional acceleratory phenomenon (RAP). We previously showed that restoration of a local bone defect in the rat leads not only to RAP but also leads to a systemic acceleration of osteogenesis (systemic acceleratory phenomenon, SAP) in distant sites of the skeleton. In this study we investigated the impact of immobilization of the defect-bearing extremity on the development of SAP. A hole 1.2 mm in diameter was drilled in the diaphysis of the left tibia of female rats. In the experimental group (n = 15), a knee tenotomy was performed in the defect-bearing left hind leg. We examined both femora, both tibiae, and the fourth lumbar vertebra by computed x-ray densitometry on day 7 postoperatively. Immobilization of the defect-bearing limb led to a decrease in x-ray density not only of the immobilized (p < 0.0001) but also of the contralateral tibia (p < 0.0001). Both femora (p < 0.001) and the fourth lumbar vertebra (p < 0.025) of the experimental group also showed a significant decrease in x-ray density. We previously showed that SAP leads to an increase in x-ray density of both femora. This increase is no longer detectable in animals after immobilization of the defect-bearing limb. Thus we conclude that immobilization interferes with SAP. This suggests the possible dependence of SAP on mechanical load. Furthermore, these data suggest a possible impact of local immobilization on the rest of the skeleton.
Mineral apposition rate in rat cortical bone: physiologic differences in different sites of the same tibia.
The mineral apposition rate (MAR) is a commonly used parameter for the characterization of bone formation and is often determined to test for experimental effects on cortical bone. We investigated whether there are physiologic variations in rat cortical MAR dependent on the side or site of measurement. In our experiment we used female rats. The animals were sacrificed on day 8, after double-fluorochrome labeling with calcein and tetracycline was performed. The MAR was calculated at 3.6, 5.4, 7.2, 9, and 10.8 mm from the epiphyseal growth plate of the lateral as well as of the medial endosteum of both right and left tibiae. We found a physiologic significant difference in the MAR between the lateral and the medial endosteal sites of the same tibia (p < 0.0001), especially near the epiphyseal growth plate. Regarding the same cortical side, there is a significant decrease (p < 0.0001) in the endosteal MAR with increasing distance from the epiphyseal growth plate. We conclude that the observed differences in endocortical MAR must be due to specific mechanical challenges. Because these differences are statistically significant, it is necessary to standardize the area of histomorphometric measurement not only with respect to the distance from the epiphyseal growth plate, but also with respect to the cortical side.
Percutaneous laser discectomy.
Percutaneous discectomy, for treatment of lumbar disc herniation, was first described in 1975 by Hijikta. Since its inception, a variety of instruments including rongeurs, augers, automated mechanical devices, and more recently, lasers have been developed to remove tissue from the herniated disc. The advantages of lasers include miniaturized, safe, and efficient energy delivery for percutaneous tissue removal. This article reviews the development of lasers for percutaneous discectomy.
Treatment of stress response during balanced anesthesia. Comparative effects of isoflurane, alfentanil, and trimethaphan.
Acute hypertensive responses during nitrous oxide-opioid-relaxant anesthesia are a common clinical problem. In adult men undergoing radical prostatectomy procedures and anesthetized with a standardized technique, we evaluated the effectiveness of alfentanil, isoflurane, and trimethaphan in treating acute hemodynamic and stress hormone responses to surgical stimulation. Stress hormone concentrations were measured 1 min before skin incision, after the onset of an acute hypertensive response, and after returning the mean arterial pressure to within 10% of the preincision values with one of the three treatment modalities. Pretreatment plasma alfentanil concentrations (151 +/- 47 to 156 +/- 47 ng.ml-1) and end-tidal nitrous oxide concentrations (66 +/- 2 to 68 +/- 2%) were similar in all three groups. Acute hypertensive events were associated with significantly increased concentrations of catecholamines and vasopressin (antidiuretic hormone [ADH]). Whereas intravenous alfentanil returned all hormone concentrations to preincision values, norepinephrine and glucose concentrations were significantly increased after adjunctive isoflurane administration. Although trimethaphan decreased the norepinephrine concentration, the epinephrine, beta-endorphin, cortisol, ADH, and glucose concentrations were significantly increased compared to preincision values. However, the persistent elevation in the posttreatment ADH concentration in the trimethaphan group was the only significant difference between the three groups. Mean (+/- standard deviation) times to awakening (2.8 +/- 3.3 to 3.8 +/- 4.2 min), extubation (8.1 +/- 4.8 to 10.3 +/- 8.5 min), and orientation (19.6 +/- 20.4 to 24.6 +/- 19.1 min) were similar in all three groups. Naloxone was required more frequently in patients in the alfentanil (35%) and isoflurane (24%) groups than in the trimethaphan group (4%).(ABSTRACT TRUNCATED AT 250 WORDS)
Tumor necrosis factor and cardiac function.
The direct effect of tumor necrosis factor (TNF), a product of activated macrophages, on myocardial performance was determined using an isolated papillary muscle technique and a modified Langendorff preparation. Papillary muscle was obtained from male adult rats 4-5 hours after they received either 100 ng/kg TNF (group A), or 100 micrograms/kg TNF (group B) or saline (control). Group B animals exhibited significantly greater peak tension development and velocity of contraction compared with controls (p less than 0.05). In group A animals these variables were not significantly different from those of the controls (p greater than 0.05). Electrophysiologic measurements revealed a significant decrease in resting membrane potential in both group A and group B animals compared with the controls (p less than 0.05). Whole hearts perfused with serum from animals treated with TNF 18-22 hours earlier exhibited significant impairment of contractility, decreased rate of systolic pressure development, and decreased rate of relaxation compared with the controls (p less than 0.05). Coronary flow and myocardial water content were similar for both groups of perfused hearts. These data suggest that tumor necrosis factor stimulates an early beneficial effect on myocardial function, which 18-22 hours later is associated with impairment of myocardial performance. This effect appears to be serum transferable.