Biomedical subjects
Publications and source records attributed to R Vogel.
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Low level chemiluminescence (CL) can be observed from autoclaved liquid culture media, as used in microbiology. The light emission is oxygen-dependent and arises from reactions following auto-oxidation of reducing Maillard products which are formed during autoclaving. The inhibition of this CL by radical scavengers and antioxidants has been studied. As superoxide radicals and hydrogen peroxide are predominantly involved in the initiation of the CL, the investigation of CL from culture media offers a convenient tool for the detection of exogenous (medium-mediated) oxidative stress being imposed onto micro-organisms in culture. Transition metal ions showed, dependent on concentration, both inhibitory and stimulating effects on the CL, which was also affected by the presence of complexing agents. Iron porphyrins and related complexes displayed a very efficient quenching of the CL, which may be of particular importance, as aerobic micro-organisms have been previously shown to be very efficient in quenching the CL from culture media.
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The glucocorticoid receptor (GR) and peroxisome proliferator-activated receptors (PPARs) play important roles in the differentiation of mesenchymal cells. Glucocorticoids acting via the GR promote osteoblastic differentiation of bone marrow stromal cells, whereas PPAR ligands induce these cells to become adipocytes. To explore potential interactions between PPAR and GR pathways in osteoblasts, we studied the interaction between PPAR subtype-selective ligands and dexamethasone (DEX) in a murine calvaria-derived osteoblastic cell line (MB 1.8) that expresses endogenous GR and PPARs. In ligand-dependent transcription assays, the PPARgamma-selective ligand TZD [(5-(4-N-methyl-N(2-pyridyl)amino)ethoxy)benzyl)thiazolidine-2,4-dione], a thiazolidinedione antidiabetic, enhanced the effect of DEX to stimulate transcription of a glucocorticoid-inducible reporter gene (mouse mammary tumor virus-luciferase). No effect was seen with PPARalpha- or hNUC1/PPARdelta-selective ligands. The GR antagonist RU-486 inhibited the DEX and TZD responses, suggesting that the effects were mediated through endogenous GR. TZD also enhanced glucocorticoid-mediated transcription in SaOS-2/B10 human osteosarcomatous cells, but not in CV-1 cells, even though both cell lines were transfected with GR plasmid and expressed significant levels of endogenous PPARgamma messenger RNA. In MB 1.8 cells, TZD decreased alkaline phosphatase activity and the expression of osteoblast-associated genes while it up-regulated the adipocyte fatty acid-binding protein. DEX counteracted the effects of TZD on alkaline phosphatase enzyme activity and osteoblastic gene expression, but enhanced the actions of TZD on adipocyte fatty acid-binding protein. Interestingly, TZD inhibited in vitro bone nodule formation and mineralization, and DEX counteracted this effect. Thus, depending on the promoter context, TZD and DEX can oppose or enhance each other's actions on gene transcription. Collectively, these results point to a complex interaction between PPAR and GR signaling pathways that regulates the effects of TZD and DEX on osteoblastic differentiation. The mechanism of this interaction is still under investigation, but might involve PPAR -dependent and -independent pathways. As thiazolidinediones represent an important new class of drugs, our findings also raise the need for further studies in bone.
Antioxidants are a heterogeneous group of agents; their mechanisms of action are diverse and are not well understood. Nonetheless, it is clear that some antioxidants, such as vitamin E, are beneficial agents in the prevention of cardiovascular disease.
The Elecsys Troponin T third generation assay uses recombinant human cardiac troponin T as standard material. The new assay has a linear calibration curve. Thus, linearity problems observed with the second generation assay have been eliminated. The assay has high precision, especially at the low end of measuring range (inter-assay CV < 10% at 0.1 microgram/L). The new standardization does not change the cut-off value of 0.1 microgram/L. The use of recombinant human cardiac troponin T as standard material enables a reproducible and reliable standardization of troponin T assays.
Pneumococcal vaccination following splenectomy is widely used as prophylaxis against overwhelming postsplenectomy infection. There remains controversy however, over the timing of vaccination. We hypothesized that delaying vaccination would increase the antibody response. Pneumococcal vaccinations were given at designated intervals to rats that had undergone either a sham abdominal surgery or splenectomy. Sixty male Sprague-Dawley rats, 250 to 400 g, were divided into three groups for vaccination: I, 1 day postoperatively; II, 7 days postoperatively; and III, 28 days postsplenectomy/sham. Serum antibody levels were then determined by enzyme-linked immunosorbent assay at 5 and 21 days after vaccination. Immunoglobulin (Ig) levels after delayed vaccination at 1 week postoperatively and 1 month postoperatively were significantly higher than levels from rats vaccinated 1 day postoperatively. IgM levels after vaccinations 1 week and 1 month postoperatively were also significantly higher than levels of rats vaccinated 1 day postoperatively (P < 0.05 for both IgG and IgM). On the basis of these results, we conclude that delaying vaccination after splenectomy enhances antibody responses.
1. A mathematical model has been developed which describes the conductive and kinetic properties of homotypic and heterotypic gap junction channels of vertebrates. 2. The model consists of two submodels connected in series. Each submodel simulates a hemichannel and consists of two conductances corresponding to a high (H) and low (L) conductance state and a switch, which simulates the voltage-dependent channel gating. 3. It has been assumed that the conductances of the high state and low state vary exponentially with the voltage across the hemichannel. 4. The parameters of the exponentials can be derived from data of heterotypic or homotypic channels. As a result, the behaviour of heterotypic channels can be predicted from homotypic channel data and vice versa. 5. The two switches of a channel are governed by the voltage drop across the respective hemichannel. The switches of a channel work independently, thus giving rise to four conformational states, i.e. HH, LH, HL and LL. 6. The computations show that the dogma of a constant conductance for homotypic channels results from the limited physiological range of transjunctional voltages (Vj) and the kinetic properties of the channel, so a new fitting procedure is presented. 7. Simulation of the kinetic properties at the multichannel level revealed current time courses which are consistent with a contingent gating. 8. The calculations have also shown that the channel state LL is rare and of short duration, and hence easy to miss experimentally. 9. The design of the model has been kept flexible. It can be easily expanded to include additional features, such as channel substates or a closed state.
Anion exchange chromatography on diethylaminoethyl cellulose was optimized to separate the cytosolic and mitochondrial isoforms of malic enzyme from rat brain. Extracts of adult rat brain and of astroglia-rich primary cultures derived from the brains of newborn rats were analyzed for their content of the two isozymes. In the case of brain tissue 45% of malic enzyme activity was due to the cytosolic isoform. In contrast, in extracts from astroglia-rich primary cultures more than 95% of the total activity was associated with the cytosolic isozyme. From these data it is concluded that the cytosolic rather than the mitochondrial isoform of malic enzyme has prominent functions in astroglial metabolism.
BACKGROUND AND PURPOSE: In dermatology high resolution ultrasonic systems proved to be valuable in following up genuine and experimental inflammatory dermatoses. The opportunities of 20 MHz ultrasonic imaging for quantitative assessment of early and late postradiation skin reactions are investigated. MATERIAL AND METHODS: Between April and November 1996, 96 high resolution ultrasound examinations of the skin in 29 patients treated for breast cancer at the University of Ulm were analyzed. Total doses between 46 and 60 Gy were applied. The time interval between the completion of radiotherapy and ultrasonic examination was < or =3 months in 18 patients and 6-135 months in 11 patients. For examinations we used a digital high resolution ultrasonic system with a ceramic 20 MHz transducer. Irradiated and non-irradiated skin were compared. RESULTS: A change of thickness and texture of the dermis depending on the time interval between the completion of radiotherapy and ultrasonic examination and on the administered radiation dose was found. There were significant differences between irradiated and non-irradiated skin regarding the dermal thickness in early (P < 0.001) as well as in late (P = 0.0018) reactions. Echogenicity of the upper and lower corium of irradiated skin decreased in early and late reaction. In upper corium the greatest reduction of signal intensity occurred in early reactions (P = 0.0001). Early reactions of the lower corium differed significantly from late changes (P = 0.001). Discrepancies between visible skin reactions described by examining physicians and ultrasonically proven changes were obvious mainly in late reactions. CONCLUSIONS: There are specific textures of early and late postradiation skin reactions in comparison to non-irradiated skin. High resolution digital 20 MHz ultrasound is non-invasive and quantitative, and in contrast to physical examination, an easy reproducible method for assessing and documenting early and late skin reaction during and after radiation therapy treatment.
To elucidate the cellular location of mitochondrial malic enzyme in brain, immunocytochemical studies were performed. For this purpose, mitochondrial malic enzyme was purified to apparent homogeneity from bovine brain and used for the immunization of rabbits. Subjecting the antiserum to affinity purification on immobilized antigen as an absorbent yielded a purified immunoreactive antibody preparation, which was characterized by probing cytosolic and mitochondrial fractions of bovine and rat brain in western blotting. As neither cross-reactivity with cytosolic malic enzyme nor immunoreactivity against other proteins could be observed, the antibody preparation was found suitable for immunocytochemistry. By using sections of perfusion-fixed rat brain, considerable resolution was achieved at the light-microscopic level. Distinct and specific staining of neurons was observed; in contrast, no staining of astrocytes and possibly unspecific staining within the nuclei of oligodendrocytes were obtained. From these data, it is concluded that mitochondrial malic enzyme is located in neurons; however, in astrocytes, the enzyme appears to be either lacking or present at a much lower level. A protective role against oxidative stress in neurons is proposed for mitochondrial malic enzyme.
Cardiovascular risk factors promote the development of cardiovascular disease by producing endothelial dysfunction. Interventions that improve endothelial function would therefore be expected to help prevent coronary disease. Studies have demonstrated that a variety of interventions, both pharmacological and nonpharmacological, can improve endothelial dysfunction. Several pharmacological interventions are briefly reviewed, with an emphasis on current issues and controversies.
There is some evidence that the incidence of acute pancreatitis is increasing worldwide. Improved treatment concepts, especially in the severe course of the disease, have significantly reduced formerly high mortality. According to the different clinical courses it is of the utmost importance for the therapeutic approach to this disease to differentiate between mild (morphologically characterized as edema) and severe (intra- and extrapancreatic necroses) as early as possible. In this respect, contrast-enhanced CT scanning and the determination of so-called necrosis indicating parameters (e.g. C-reactive protein) have been established as the "gold-standard". While patients with acute edematous pancreatitis are successfully treated in a normal ward, patients with a proven necrotizing course of the disease should undergo intensive monitoring and maximum intensive care therapy in the ICU. Additionally, these latter patients should receive antibiotics which are capable of penetrating the pancreas and the pancreatic necroses in bactericidal concentrations. It seems more and more evident that only patients under this treatment regimen who develop infected pancreatic necrosis and sepsis are candidates for surgical intervention. Infected pancreatic necrosis can be easily diagnosed with a high level of safety and reliability by fine needle puncture and aspiration of pancreatic necrosis and fluid collections under imaging-guided procedures. Patients with sterile necrosis respond in most cases to intensive care therapy and in these patients the indication for surgery will be only exceptional. Surgery should be performed as late as possible to ensure sufficient demarcation of the necroses. In our experience the best surgical treatment modality for infected pancreatic necrosis is necrosectomy combined with postoperative continuous local lavage of the retroperitoneum. Mortality of severe acute pancreatitis has been reduced under this treatment concept to below 10%.
1. Cell pairs of the insect cell line Sf9 (Spodoptera frugiperda) were chosen to examine the electrical properties of gap junction channels. The dual voltage-clamp method was used to control the membrane potential of each cell (Vm,1 and Vm,2) and hence the junctional voltage gradient (Vj), and to measure intercellular current. 2. Studies with preformed pairs revealed that the gap junction conductance (gj) is controlled by a Vj- and a Vm-sensitive gate. At steady state, gj = f(Vj) was bell shaped and symmetrical (Boltzmann: Vj.0 = -54 and 55 mV, the normalized minimum conductance at large Vj values (gj,min) = 0.24 and 0.23, z = 5.5 and 6.1 for negative and positive Vj, respectively) and gj = f(Vm) was S shaped (Vm.0 = 13 mV, gj,min = 0, z = 1.5). 3. Single channels exhibited two conductances, a main state (gamma j,main) of 224 pS and a residual state (gamma j,residual) of 42 pS. 4. We conclude that gap junctions in Sf9 cells behave similarly to those in the insect cell line C6/36 (Aedes albopictus). 5. An induced cell pair approach was used to examine heterotypic gap junction channels between Sf9 cells and C3/36 cells. 6. Heterotypic channels showed a gamma j,main of 303 pS and a gamma j,residual of 45 and 64 pS, depending on whether the Sf9 cell or C6/36 cell was positive inside. 7. In heterotypic gap junctions, gj = f(Vj) was bell shaped and asymmetrical (gj was more sensitive to Vj when the C6/36 cell was positive inside) and gj = f(Vm) was S shaped (Vm,0 = 2 mV, gj,min = 0, z = 2.9). 8. We conclude that heterotypic channels possess a Vj- and Vm-sensitive gating mechanism. Vj gating involves two gates, one located in each hemi-channel. Vj gates are operated independently and close when the cytoplasmic aspect is made positive. 9. A comparison of homo- and heterotypic channel data suggests that docking of hemi-channels may affect channel gating, but not channel conductance.
This study examined the optimal angiographic definition for long-term success after angioplasty and compared visual and quantitative angiographic measurements in assessing outcome. The National Heart, Lung, and Blood Institutes--Percutaneous Transluminal Coronary Angioplasty Registry prospectively followed 1,768 patients from 15 clinical centers. Symptom-free survival, defined as survival without angina, myocardial infarction, bypass surgery, or death, occurred in 59% of patients. In a subset of 393 patients, quantitative coronary angiography (QCA), done at a core angiographic laboratory, was compared with visual site readings. Although there was considerably more variability for visual readings, a site reading of a change in percent stenosis of >20% correlated highly with symptom-free survival (64.6% for patients who had all lesions successfully dilated, 48% for patients with partial success, and only 21% for patients without angiographic success; p < 0.001). Similar findings were seen for other angiographic definitions, but a change of > 20% was most discriminatory. In contrast, QCA readings had little or no predictive value. This study confirms that visual assessment of the immediate change in percent stenosis is predictive of a successful 1-year outcome. A change of greater than 20% is most discriminatory and should still be used to define angiographic success. QCA does not appear to be superior to visual assessment in predicting 1-year outcome.
Glucagon-like peptide 1 (GLP-1) is an insulinotropic hormone released after nutrient ingestion which is known to augment insulin secretion, inhibit glucagon release, and promote insulin-independent glucose disposition. To determine the overall effect of GLP-1 on glucose disposition after a meal we studied a group of healthy, conscious baboons before and after intragastric glucose administration during infusions of saline, and two treatments to eliminate the action of GLP-1: (a) exendin-[9-39] (Ex-9), a peptide receptor antagonist of GLP-1; or (b) an anti-GLP-1 mAb. Fasting concentrations of glucose were higher during infusion of Ex-9 than during saline (4.44 +/- 0.05 vs. 4.16 +/- 0.05 mM, P < 0.01), coincident with an elevation in the levels of circulating glucagon (96 +/- 10 vs. 59 +/- 3 ng/liter, P < 0.02). The postprandial glycemic excursions during administration of Ex-9 and mAb were greater than during the control studies (Ex-9 13.7 +/- 2.0 vs. saline 10.0 +/- 0.8 mM, P = 0.07; and mAb 13.6 +/- 1.2 vs. saline 10.6 +/- 0.9 mM, P = 0.044). The increments in insulin levels throughout the absorption of the glucose meal were not different for the experimental and control conditions, but the insulin response in the first 30 min after the glucose meal was diminished significantly during treatment with Ex-9 (Ex-9 761 +/- 139 vs. saline 1,089 +/- 166 pM, P = 0.044) and was delayed in three of the four animals given the neutralizing antibody (mAb 946 +/- 262 vs. saline 1,146 +/- 340 pM). Thus, elimination of the action of GLP-1 impaired the disposition of an intragastric glucose meal and this was at least partly attributable to diminished early insulin release. In addition to these postprandial effects, the concurrent elevation in fasting glucose and glucagon during GLP-1 antagonism suggests that GLP-1 may have a tonic inhibitory effect on glucagon output. These findings demonstrate the important role of GLP-1 in the assimilation of glucose absorbed from the gut.
Two forms of the transmembrane human protein tyrosine phosphatase (PTP sigma), generated by alternative splicing, were identified by cDNA cloning and Northern hybridization with selective cDNA probes. The larger form of PTP sigma is expressed in various human tissues, human osteosarcoma, and rat tibia. The hPTP sigma cDNA codes for a protein of 1911 amino acid residues and is composed of a cytoplasmic region with two PTP domains and an extracellular region that can be organized into three tandem repeats of immunoglobulin-like domains and eight tandem repeats of fibronectin type III-like domains. In the brain, the major transcript of PTP sigma is an alternatively spliced mRNA, in which the coding region for the fibronectin type III-like domains number four to seven are spliced out, thus coding for a protein of 1502 amino acid residues similar to the rat PTP sigma and rat PTP-NE3. Using in situ hybridization, we assigned hPTP sigma to chromosome 6, arm 6q and band 6q15. The bacterial-expressed hPTP sigma exhibits PTPase activity that was inhibited by orthovanadate (IC50 = 0.02 microM) and by two bisphosphonates used for the treatment of bone diseases, alendronate (ALN) (IC50 = 0.5 microM) and etidronate (IC50 = 0.2 microM). In quiescent calvaria osteoblasts, micromolar concentrations of vanadate, ALN and etidronate stimulate cellular proliferation. These findings show tissue-specific alternative splicing of PTP sigma and suggest that PTPs are putative targets of bisphosphonate action.