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Biomedical subjects

R Roth

Publications and source records attributed to R Roth.

At least 91 records · Page 5Linked to original sources

Characterization and partial purification of an ATPase and inorganic pyrophosphatase of the archaebacterium Methanobacterium thermoautotrophicum.

ATPase and inorganic pyrophosphatase (PPase) activities have been detected in several methanogenic bacteria. These activities are believed to play a crucial role in energy metabolism. In the present study we have investigated some characteristics of the ATPase and the inorganic PPase activities of Methanobacterium thermoautotrophicum. Although these proteins migrate identically on non-dissociating gels, they are catalyzed by distinct enzymes which are separable by biochemical purification methods. The partially purified enzymes are composed of at least two subunits. The ATPase subunits have molecular masses of about 43 and 33 kDa and the inorganic PPase such of about 31 and 25 kDa. After purification, the PPase and the ATPase did not hydrolyze ATP and PP(i), respectively. The membrane-bound ATPase and PPase activities are distinguished in response to sodium fluoride, by the effects of divalent cations, by the temperature ranges for activities and the solubilization behaviour by different extractants. Most investigated catalytic and structural properties of the ATPase do not suit the current criteria for classifying this enzyme under either the F-, V- or P-ATPases.

Adenosine Triphosphatases↗

Golgi membrane dynamics imaged by freeze-etch electron microscopy: views of different membrane coatings involved in tubulation versus vesiculation.

We used high resolution three-dimensional electron microscopy to visualize Golgi cisternal structure and analyze morphological transitions induced by various in vitro incubations. Our images show that Golgi cisternae have two distinct surface coatings with different distributions and apparent functions. The first type, probably a coatomer coat, consists of tightly packed approximately 10 nm surface particles. These are localized exclusively to sites of membrane budding and are as intimately involved in bud formation as clathrin or caveolin coats. When this coating is exaggerated by GTP gamma S, the periphery of all cisternae is partitioned into coated vesicles that remain attached at their sites of formation. A second, much finer coating is evenly distributed over the periphery of cisternae, including tubules enclosing the fenestrae. It appears to stabilize the membrane curvature associated with tubules and edges. These different coatings must be considered in further attempts to unravel Golgi membrane trafficking mechanisms.

Animals↗

Liver hemodynamics during portal venous endotoxemia in swine.

The acute hemodynamic response of the liver to portal endotoxemia was measured in six isoflurane anesthetized pigs in which volume support was used to maintain normal cardiac output. After baseline monitoring, bacterial endotoxin (LPS) was infused over 1 hr into a mesenteric vein at a rate of 1 microgram.kg-1.hr-1, and monitoring was continued for 1 hr postinfusion. Peak vasoconstriction occurred during LPS infusion in both the hepatic artery (resistance increases 349% of baseline, P < 0.05) and the liver's portal circulation (resistance increases 159% of baseline, P < 0.05). Increased vascular resistance was also detected in lung (increases 433% of baseline) and intestine (increases 130% of baseline) at the midpoint of the LPS infusion. The non-splanchnic circulation, defined for our analysis as all of the peripheral circulation except the portal and hepatic arterial circulation, generally exhibited little change in vascular resistance during LPS infusion. LPS was incompletely cleared by the liver, but secondary clearance by the lung prevented large increases in the LPS concentration of arterial blood. During the first hour postinfusion, the systemic vascular resistance subsequently decreased to near normal in all vascular beds, with the exception of the liver's portal circulation. A sustained and secondary increase in vascular resistance of the liver's portal circulation and portal vein pressure occurred during the first hour after LPS infusion. We conclude that most of the vasoconstriction in the acute response to portal endotoxemia occurs in the liver and lung, organs directly exposed to elevated levels of endotoxins.

Animals↗

[Radiation sensitivity of the normal bile duct during high dose rate afterloading irradiation with Iridium 192. Experimental studies in pigs].

This animal study was designed to assess the tolerance of the normal bile duct to a single intraluminal high-dose-rate afterloading irradiation. This information is essential for treatment recommendations in irradiation therapy of malignant bile duct stenoses. In 16 pigs bile duct catheters were inserted surgically. Over these devices single intraductal doses of 7.5 Gy and 15 Gy were applied using an iridium-192 source. After a period of two to three months the bile ducts and the surrounding structures were investigated by means of cholangiography, angiography, and macropathological and histological investigations. A single intraductal dose of 7.5 Gy leads to a slightly scarred shrinkage of the bile duct with sclerosis and obliteration of the capillary vessels. Vasculitis and necrosis of the bile duct wall are still evident three months after irradiation. A single dose of 15 Gy leads to severe lesions with widespread necroses of the bile duct wall which tend to bleed, and result in a high degree of shrinkage of the bile duct. The radiation damage is most prevalent near the papilla and in the narrow segments near the liver. Single intraluminal high-dose-rate afterloading doses of 7.5 and 15 Gy cause significant lesions and complications at the bile ducts. As the intact bile duct is the Achilles heel of intraductal therapy, considerably lower single doses are recommended for a fractioned treatment.

Animals↗

[The long-term therapy of familial hypercholesterolemia with heparin-induced extracorporeal LDL precipitation].

The long-term tolerance to and effectiveness of heparin-induced extracorporeal LDL precipitation (HELP) in combination with lipid reducing drugs and diet was tested in six patient (5 males, 1 female; mean age 48 +/- 4 years). Follow-up period was over 50 months, in one patient over 24 months, while one man had a sudden cardiac death 57 weeks after starting treatment. The study was divided into three phases. In phase I (24 months) treatment consisted of HELP and conventional lipid-reducing drugs; in phase II (12 months) of lovastatin (80 mg daily) and cholestyramine (12-24 g daily); and phase III (14 months) of HELP, lovastatin and cholestyramine. In phase I it was possible to lower the pre-treatment level of LDL-cholesterol from 306 +/- 18 mg/dl to 173 +/- 13 mg/dl (43.5%). A similar effect (from 307 +/- 21 mg/dl to 155 +/- 17 mg/dl [-49.5%]) was obtained in phase II. The resumption of HELP reduced the pre-treatment LDL concentration to 136 +/- 9 mg/dl (-55.7%). The various treatment regimens were well tolerated. Biochemical data remained unchanged except for iron loss requiring substitution. Thus combined HELP, lovastatin and ion exchange offer for the first time an effective and reliable means in familial hypercholesterolaemia of clearly reducing long-term the mean LDL cholesterol level below the atherosclerosis threshold of 120 mg/dl.

Anticholesteremic Agents↗

Initial experience with a new compression device for hemostasis after femoral arterial puncture.

We evaluated a new hemostasis device to maintain focused pressure over the femoral artery puncture site for 2-6 hours in 330 cases after diagnostic catheterization or coronary angioplasty. No patient developed later hematoma or vascular complications. The initial favorable experience suggests that this hemostasis device has advantageous features of stability and site visualization and is a suitable adjunct to current methods of maintaining puncture site pressure after brief manual compression following arterial catheterization.

Cardiac Catheterization↗

Factors which provoke post-infarction depression: results from the post-infarction late potential study (PILP).

Symptoms of depression in the majority of patients immediately following acute myocardial infarctions (AMI) resolve rapidly; they are an adjustment reaction. However, in a group of 552 male patients there were 80 (14.5%) patients with persistent major depressive symptoms during a finite period after AMI. Infarction size was assessed by maximum creatine kinase levels, the QRS-complex and the occurrence of late potentials. These measures did not correlate with the degree of depressed moods in these groups. An arrhythmic event in the early hospitalization phase, a recurrent infarction, dyspnoea, and persistent angina pectoris before the AMI were significantly related to more profound degrees of depression. Patients who reported serious life-events in the last 2 yr before AMI, or who suffered from exhaustion and fatigue in the prehospital phase were subject to significantly higher levels of depression. A prodromal phase prior to hospitalization free of bodily symptoms and the use of denial were related to low levels of depression. The logistic regression model incorporating all univariate significant variables revealed that symptoms of exhaustion and fatigue prior to AMI had the strongest independent correlation with post AMI depression.

Activities of Daily Living↗

Influence of radiographic contrast media viscosity to flow through coronary angiographic catheters.

To assess the influence of viscosity on flow resistance, 4 clinically available contrast media were injected through 12 angiographic catheters of varying dimensions at 20 degrees and 37 degrees C. Seven cc of contrast was injected at a pressure of 200 PSI at 3cc/sec by Medrad Mark IV power injector. The pressure of injection through the manifold was recorded with an electronic pressure transducer. The lowest injection pressure at 37 degrees C occurred with Hexabrix. Differences in contrast media viscosity were apparent with catheters less than 6 French diameter. There were minimal differences in injection pressures with regard to the coronary curve tip configurations for any of the contrast agents. At 20 degrees C, Isovue had lower injection pressures than the other contrast agents. Injection through 5 French catheters demonstrated a greater than 1.5 atmosphere difference, especially between Omnipaque and Hexabrix. The difference in contrast media injection pressure was greater than 2 atmospheres between 8 French guiding and 8 French diagnostic catheters and between 5 French and 6 French diagnostic catheters and less than 2 atmospheres between 8 French and 6 French diagnostic catheters. Injection pressure differences greater than 1 atmosphere were not observed for catheters of the same French size at body or room temperature contrast injection. These data indicate that important temperature related viscosity differences between agents are present and confirm that the largest differences in contrast media are most apparent for the smallest diameter catheters. Given equivalent image opacification and hemodynamic and adverse effects, selection of a low viscosity contrast media theoretically provides an advantage during procedures using small diameter catheters or interventional procedures requiring contrast media visualization through reduced channels.

Angiography↗

Characterization of latent recombinant TGF-beta 2 produced by Chinese hamster ovary cells.

Latent recombinant transforming growth factor-beta 2 (LrTGF-beta 2) complex has been purified from serum-free media conditioned by Chinese hamster ovary cells transfected with a plasmid encoding the TGF-beta 2 (414) precursor. Under neutral conditions, LrTGF-beta 2 had an apparent molecular weight of 130 kDa. The complex contained both mature and pro-region sequences. Acidification of LrTGF-beta 2 resulted in the release of mature 24 kDa TGF-beta 2 from the high molecular weight pro-region-containing complex, suggesting that TGF-beta 2 was non-covalently associated with this complex. These results were confirmed by crosslinking experiments performed on partially purified LrTGF-beta 2. Protein sequence analysis of the purified TGF-beta 2 pro-region indicated that signal peptide cleavage occurred between ser(20) and leu(21). The pro-region, which previously was found to contain mannose-6-phosphate, bound to the mannose-6-phosphate receptor. Proteolytic cleavage of mature TGF-beta 2 from pro-TGF-beta 2 was inhibited by monensin and chloroquine suggesting that binding to this receptor and subsequent transport to acidic vesicles may be involved in the processing of rTGF-beta 2 precursor.

Amino Acid Sequence↗

[Patient-specific determinants of delay in goal-oriented patient behavior in acute myocardial infarct. Results of the post-infarct late potential study].

369 (63%) of 586 consecutive patients with confirmed myocardial infarction were admitted to hospital within 6 h of symptom onset. Patients' arrival to hospital followed a characteristic circadian distribution with a marked morning increase between 0600 and 1200 hours, and a corresponding decrease in the late night hours. There were no differences in the occurrence of late potentials and in ECG data between early and late arrival group. The early group (less than 5.9 h) was, however, characterized by significantly higher cardiac enzyme levels than the late group (greater than 6.0 h); they also experienced more arrhythmic events in the acute phase (16.0% vs. 9.7%; p less than 0.032). Catecholamines were significantly more often necessary (26.4% vs. 10.3%; p less than 0.0001). Six-month prognosis after AMI was markedly worse. The time between symptom onset and hospital arrival was not affected by age and risk factors. In univariate analysis, pain history and recurrent infarction also had no influence on delay of admission to hospital. The time interval was, however, significantly shorter when the acute event occurred during the night (18.4% vs. 8.3%; p less than 0.0001). Absence of prodromi in the prehospital phase (18.5% vs. 12.0%; p less than 0.04) and a higher socio-economic level (68.3% vs. 61.0%; p less than 0.077) also shortened the admission time, whereas a hyperactive behavioral pattern prolonged the delay time (21.1% vs. 28.9%; p less than 0.05).

Adult↗

Effects of heparin treatment on colonic torsion-associated hemodynamic and plasma eicosanoid changes in anesthetized ponies.

Large colon torsion frequently is a fatal condition in horses. The purpose of the study reported here was to determine systemic arterial pressure, plasma eicosanoid concentrations, colonic blood flow, vascular resistance, tissue pH, and morphologic features associated with large colon torsion and detorsion, and to evaluate the effects of sodium heparin (80 IU/kg of body weight, IV) treatment on these values. Values were determined in 20 anesthetized ponies that were randomly assigned into 4 equal groups: control; control/heparin; torsion; torsion/heparin. Torsions were created by a 720 degrees rotation of the cecum and colon around their long axes at the sternal and diaphragmatic flexures. After 1 hour of torsion, the torsion was corrected and the colon was allowed to reperfuse for 1 hour. Heparin was administered 30 minutes into the experiment. Parametric data were analyzed (P less than or equal to 0.05), using split-plot analysis of variance, with differences between means evaluated with a modified Bonferroni t test; histopathologic data were analyzed (P less than or equal to 0.05) with a Kruskal-Wallis one-way analysis of variance by ranks. Heparin prevented colonic detorsion-induced hypotension and increases in vascular resistance and thromboxane concentration, and it significantly increased colonic blood flow for 40 minutes during reperfusion. Heparin did not alter prostacyclin concentration or the histologic appearance of the large colon.

Anesthesia↗