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S Witt

Publications and source records attributed to S Witt.

At least 37 records · Page 2Linked to original sources
PubMed1997

Differential regulation of mitogen-activated protein/ERK kinase (MEK)1 and MEK2 and activation by a Ras-independent mechanism.

Mitogen-activated protein (MAP)/ERK kinase (MEK)1 and MEK2 are the upstream activators of the MAP kinases, ERK1 and ERK2. MEK1 and MEK2 are approximately 85% identical in sequence but have unique inserts in their C-terminal domains. MEK isoform-specific antibodies were used to examine expression and regulation of each enzyme. MEK1 and MEK2 were expressed in approximately equal amounts in several cell lines; in some, MEK1 was present in slight excess. Activation of tyrosine kinase-containing receptors, heterotrimeric G proteins, and protein kinase C enhanced the activities of both MEK isoforms in 293 and PC12 cells. AIF4-stimulated both MEK1 and MEK2 in PC12 cells expressing a dominant interfering Ras mutant that prevents nerve growth factor-dependent activation of the cascade. Carbachol also stimulated the pathway in these cells. Thus, in addition to their ability to activate Ras/Raf and the downstream ERK pathway, heterotrimeric G proteins also appear to trigger a Ras-independent mechanism to regulate this kinase cascade. In U373, Chinese hamster ovary (CHO), and INS-1 cells, MEK1 was activated by regulators of ERKs, while MEK2 was not. These data suggest that, like the MAP kinases ERK1 and ERK2, in some cell settings the two similar MEK isoforms are differentially regulated.

3T3 Cells↗
PubMed1996

Correlates of the hemodynamic determinants of blood pressure.

The purpose of this study was to investigate the association of sex, race, lean body mass, and fat mass with the hemodynamic determinants of blood pressure, including stroke volume, heart rate, and total peripheral vascular resistance. The study included 201 subjects aged 6 to 17 years, 105 of whom were male and 98 of whom were black. Lean body mass and fat mass were both significant (P < .05) independent determinants of stroke volume, cardiac output, and total peripheral vascular resistance. However, the direction of the effect of lean body mass was opposite for stroke volume and cardiac output compared with that of total peripheral vascular resistance. The direct relationship of lean body mass with systolic blood pressure (SBP) and diastolic blood pressure (DBP) (regression coefficients, 0.55 +/- 0.05 for SBP and 0.47 +/- 0.05 for DBP) indicates that the effect of lean body mass on cardiac output may predominate. Lean body mass explained substantially more of the variance of the hemodynamic variables than did fat mass. After control for the effects of body size, male subjects had higher heart rate and cardiac output, and female subjects had higher vascular resistance. White subjects had higher stroke volume and cardiac output, and black subjects had higher peripheral vascular resistance. This study demonstrates that lean body mass is a more important correlate of the hemodynamic determinants of blood pressure than is fat mass and that sex and race have significant independent relationships with the hemodynamic determinants of blood pressure in children and adolescents.

Adolescent↗
PubMed1996

Initial evaluation of an interactive test of sentence gist recognition.

The laser videodisc-based Sentence Gist Recognition (SGR) test consists of sets of topically related sentences that are cued by short film clips. Clients respond to test items by selecting picture illustrations and may interact with the talker by using repair strategies when they do not recognize a test item. The two experiments, involving 40 and 35 adult subjects, respectively, indicated that the SGR may better predict subjective measures of speechreading and listening performance than more traditional audiologic sentence and nonsense syllable tests. Data from cochlear implant users indicated that the SGR accounted for a greater percentage of the variance for selected items of the Communication Profile for the Hearing-Impaired and the Speechreading Questionnaire for Cochlear-Implant Users than two other audiologic tests. As in previous work, subjects were most apt to ask the talker to repeat an utterance that they did not recognize than to ask the talker to restructure it. It is suggested that the SGR may reflect the interactive nature of conversation and provide a simulated real-world listening and/or speechreading task. The principles underlaying this test are consistent with the development of other computer technologies and concepts, such as compact discinteractive and virtual reality.

Adult↗
PubMed1995

Effect of lean body mass, fat mass, blood pressure, and sexual maturation on left ventricular mass in children and adolescents. Statistical, biological, and clinical significance.

BACKGROUND: Left ventricular hypertrophy has been established as an independent risk factor for the development of cardiovascular morbidity and mortality. It is clear that left ventricular mass increases during childhood and adolescence with body growth. The extent to which other factors, such as obesity, stage of sexual maturation, and level of blood pressure, determine left ventricular mass has been controversial. METHODS AND RESULTS: The study was a cross-sectional evaluation of the relationship of left ventricular mass determined by echocardiography with lean body mass and fat mass determined by dual-energy x-ray absorptiometry, which is the most valid and reliable method for determination of body composition in children and adolescents. The relationship of left ventricular mass with the stage of sexual maturation and with systolic and diastolic blood pressure was also evaluated. Two hundred one subjects (105 boys, 96 girls; 103 white and 98 black) 6 to 17 years old were studied. Age (r = .72), height (r = .81), weight (r = .84), body surface area (r = .87), sexual maturation (r = .75), lean body mass (r = .86), fat mass (r = .54), systolic BP (r = .58), and diastolic BP (r = .48) were all univariate correlates of left ventricular mass. In a multiple regression analysis, only lean body mass, fat mass, and systolic blood pressure were statistically significant independent correlates of left ventricular mass. Lean body mass alone explained 75% of the variance of left ventricular mass, whereas fat mass and systolic blood pressure explained only 1.5% and 0.5% of the variance, respectively. Lean body mass was the strongest determinant of left ventricular mass in all four race-sex groups. CONCLUSIONS: This study provides an opportunity to separate the effects on left ventricular mass of lean body mass resulting from linear growth from those of fat mass resulting from obesity. Lean body mass, fat mass, and systolic blood pressure all have a statistically significant independent association with left ventricular mass, suggesting that all three play an important biological role in determining left ventricular mass. However, fat mass and systolic blood pressure have only a small impact on left ventricular mass. This indicates that fat mass and blood pressure would be expected to be of only minor clinical importance in determining left ventricular mass in normal children and adolescents.

Absorptiometry, Photon↗
PubMed1995

Effects of talker familiarity on communication breakdown in conversations with adult cochlear-implant users.

This investigation had three objectives: a) to determine the types of repair strategies that cochlear-implant users implement to rectify communication breakdowns during ongoing conversation when talking to either familiar or unfamiliar communication partners, b) to determine how communication partners respond to particular types of repair strategies, and c) to describe the use of conversational behaviors that might circumvent communication difficulties. In Experiment 1, cochlear-implant subjects were videotaped while talking to familiar and then unfamiliar communication partners. In Experiment 2, a second group of cochlear-implant subjects were videotaped while speaking with an unfamiliar partner for 6.5 minutes. Analysis of the videotapes revealed that the cochlear-implant subjects in the two experiments most commonly asked "what?," "huh," or "pardon?" after not recognizing a spoken message (e.g., following a communication breakdown), regardless of whether the communication partner was familiar or unfamiliar. Communication partners' more common response to this repair strategy was to repeat the original message. When cochlear-implant subjects repeated back the segment of a message that they understood, communication partners most often confirmed or corrected them. When they requested information, communication partners usually provided it. The cochlear-implant subjects were more likely to use controlling conversational behaviors when interacting with unfamiliar than familiar communication partners. We conclude that repair strategy-response adjacency pairs may emerge during spontaneous conversations. Use of both specific and nonspecific repair strategies may indicate cochlear-implant users' adherence to a cooperative principle.

Adult↗
PubMed1994

Pressure-time product and work of breathing during biphasic continuous positive airway pressure and assisted spontaneous breathing.

The pressure-time product (PTP) and work of breathing (W) were measured in 19 intubated patients during weaning from mechanical ventilation after aortocoronary bypass surgery. The patients were supported by two different ventilatory modes: biphasic continuous positive airway pressure (Biphasic CPAP; a ventilatory mode that permits spontaneous breathing at two different levels of airway pressure during inspiration and during expiration) and pressure-support ventilation (denominated as assisted spontaneous breathing, ASB). Our aim was to compare the energy cost of breathing imposed by both modes. The PTP was obtained by integrating the area enclosed by the esophageal and chest-wall static recoil pressure curves. The W performed by the patient (Wp) was calculated using Campbell's diagram method, based on the esophageal pressure curve and normalized per liter of ventilation. Two comparable support levels, corresponding to a high and a low degree of mechanical support, were studied for each ventilator mode. The levels of support were generally higher than that necessary to overcome inspiratory resistance. We found significantly higher PTP values during biphasic CPAP than during ASB (p < 0.01), whereas the Wp was the same with biphasic CPAP and ASB at both support levels. We conclude that because of the higher PTP values measured during biphasic CPAP, this mode of ventilation appears to have been more exhausting for our patients. The discrepancy between PTP and Wp is probably due to the different mechanisms of support given by the two ventilatory modes and the greater patient effort is related to those respiratory cycles that are not helped by the ventilator and which represent nearly half of the breaths during biphasic.

Aged↗
PubMed1993

Characterization of monoclonal islet cell reactive autoantibodies from the diabetic biobreeding (BB/OK) rat.

Two fusion experiments using the heteromyeloma cell line CB-F7 and splenocytes from two diabetes-prone BB (BioBreeding) rats at the onset of diabetes resulted in 128 islet cell reactive autoantibodies primarily detected with permeabilized insulin-producing rat insulinoma cells (RIN) by a cellular enzyme-linked immunosorbent assay. Seventy-nine (62%) of 128 RIN cell reactive supernatants exhibited a cross-reactivity with rat splenic lymphocytes. Six stable hybridomas secreting monoclonal ICSA (islet cell surface antibodies) were established, but only one monoclonal antibody, R4B10, showed preferential beta-cell binding. Six monoclonal antibodies showed a dual reactivity as ICA (islet cell cytoplasmic antibodies) detected by immunostaining of pancreatic islet cryosections and as ICSA on the surface of viable islet cells, whereas two reacted only with an ICA-like pattern. One monoclonal ICSA was specifically displaced from the RIN cell surface by sera of type 1 diabetic patients.

Animals↗
PubMed1992

Monoclonal antibody-mediated cytotoxicity against rat beta cells detected in vitro does not cause beta-cell destruction in vivo.

Two monoclonal Beta-cell surface antibodies M10H6 und K14D10 were obtained by fusion of spleen cells of Balb/c mice with the myeloma cell line P(3)0. The monoclonal antibody M10H6 was induced by immunization with rat insulinoma cells finally boostered with disintegrated rat islets, whereas the K14D10 was generated after immunization with porcine proinsulin. Both monoclonals belong to the IgG2A isotype and were screened with insulin-producing rat insulinoma cells by an indirect immunofluorescence test as well as by a cellular enzyme linked immunosorbent assay. In addition to the cell surface binding on living Beta cells the monoclonals react with islets on cryostat sections of rat pancreas. The anti-islet cytotoxic potential of these monoclonals was measured by 51Chromium-release in the presence of complement or Fc-receptor bearing leucocytes using 51Chromium-labelled rat islet cells as target. Both antibody secreting hybridomas were propagated in syngeneic mice resulting in high levels of islet cell surface antibodies in ascites and sera from the recipient. High anti-islet cytotoxicity was mediated by ascites fluid, but no mouse developed hyperglycaemia. Furthermore, the repeated injections of the monoclonals into rats did not exert a diabetogenic action and failed to reduce the pancreatic insulin content although the attraction of the K14D10 to the pancreatic islets in vivo could be demonstrated. We conclude that islet cell surface antibody-mediated Beta-cell lysis in vitro may not be relevant to Beta-cell destruction in vivo.

Animals↗
PubMed1991

Influence of target cell preparation on binding of monoclonal islet cell reactive antibodies (mc-ICRA) in cellular enzyme-linked immunosorbent assay (CELISA).

A rapid, effective and sensitive CELISA for the detection of monoclonal islet cell reactive antibodies (mc-ICRA) using the insulin-producing rat insulinoma cell line (RIN) is described. RIN cells are a suitable target for this monoclonal antibody assay as shown by a comparative study with normal rat islet cells. We tested the influence of the target cell preparation and obtained the best sensitivity and reliability with the CELISA using desiccated cells or desiccated cell homogenate with a cell number of 5 x 10(4) cells per well rather than an adsorbed cell homogenate. Furthermore, ethanol fixation of RIN cells resulted in a loss of antigenicity as shown particularly by the detection of islet cell surface antibodies. We also compared the binding of mc-ICRA in RIN-CELISA with data obtained by indirect immunofluorescence using viable RIN cells as targets. By permeabilization of the cell membrane by desiccation or sonication, more antibodies are detected in CELISA (surface and cytoplasmic antibodies), whereas in immunofluorescence on viable RIN cells, only surface reactive antibodies are detected.

Animals↗
PubMed1991

Central antiaversive and antinociceptive effects of anterior pretectal nucleus stimulation: attenuation of autonomic and aversive effects of medial hypothalamic stimulation.

Previous studies have shown that stimulation of the rat anterior pretectal nucleus (APtN) strongly depresses a spinal reflex to noxious heat without causing significant aversion or depression of other motor responses. It is not known if APtN stimulation can similarly reduce the aversiveness of electrical stimulation of the brain, nor is it known if APtN stimulation is itself rewarding or aversive. This study used a simple switch-off paradigm to examine the rewarding properties of APtN stimulation at different sites throughout the nucleus and also used the tail-flick test to determine if the stimulation produced antinociception. The effects of APtN stimulation on the behavioural and autonomic responses to electrical stimulation of the medial hypothalamus (MH) and the nucleus raphe magnus (NRM) were also examined. The results show that electrical stimulation of dorsal APtN was rewarding and also caused antinociception which lasted for 50 min. However, sites which gave the strongest reward were not necessarily those which gave the greatest antinociception, as these effects were not correlated. Electrical stimulation of ventral APtN induced only aversive effects. The aversive and autonomic effects of MH stimulation were significantly reduced by conditioning stimulation of dorsal APtN. However, the very similar escape and autonomic effects of NRM stimulation were unaffected by APtN stimulation. These results suggest that electrical stimulation of the dorsal parts of the APtN has positive rewarding properties as well as the well-known antinociceptive effects. The antiaversive effects of dorsal APtN stimulation may be due in part to the inhibition of central substrates of aversion as well as inhibition of sensory neurones.

Animals↗
PubMed1991

[Knee joint hemarthrosis. An absolute indication for operation?].

The degree of severity of an injury of the knee joint is estimated mainly on the basis of the history given by the patient, stability testing and hemarthrosis. Once these are known the surgeon usually decides whether an arthroscopy should be done or not. In a group of 365 patients who had undergone arthroscopy after acute knee injury, we made a retrospective check of the indications. The purpose of the study was to evaluate the sensitivity of hemarthrosis as an indicator of severe injury of the knee joint necessitating operative treatment. For stability testing we used the varus and valgus stress test, the Lachman test, the Anterior drawer test, the pivot shift test and the anterior drawer test with medial and lateral rotation. The Lachman test was repeated under general anesthesia in the majority of patients just before the arthroscopy. The results were evaluated retrospectively with the aid of electronic data processing. In nearly 70% of cases the arthroscopy was indicated because of positive signs of instability together with a typical history provided by the patient or hemarthrosis. In another 27%, the hemarthrosis was the sole reason for the surgeon's decision to perform an arthroscopic investigation of the injured knee joint. In 80.5% of all cases the clinical diagnosis of lesion of the anterior cruciate ligament (ACL) or combined injury to ligaments and menisci was confirmed by the arthroscopy. In addition to this group, in another 10.8% we found other severe lesions necessitating by operation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1991

[Production of monoclonal antibodies in serum-free medium in dialysis tubing].

A simple procedure for the production of pure monoclonal antibodies (mab) in dialysis tubing has been used. Hybridomas which produce pancreatic islet cell reactive monoclonal antibodies were grown in dialysis tubing containing serum free medium. The dialysis tubing was inserted into a flask with medium containing 7.5% foetal calf serum. The flask was placed on a roller and medium was changed every two days. The optimal time for harvesting the mab could be shown to be after 10 days of culture with a 4 fold increased immunoglobulin concentration in comparison to a conventional hybridoma culture. Immunoglobulin concentrations up to 110 mg/l and cell yields of 1.3 x 10(6)/ml have been obtained. The low concentration of contaminating low molecular weight proteins in the supernatant facilitated or saved purification of mab.

Animals↗
PubMed1991

CELISA for rapid screening of monoclonal islet cell surface antibodies using living rat insulinoma cells as target.

An improved rapid cell enzyme-linked immunosorbent assay (CELISA) is described which is suitable for the large scale screening of monoclonal antibodies to islet cell surface antigens. 5 x 10(4) insulin-producing rat insulinoma (RIN) cells were seeded per well in a 96-well flat-bottomed polystyrene plate coated one day before a 0.01% poly-D-lysine solution in PBS. After culture for 4 days in 200 microliters/well RPMI 1640 supplemented with 7.5% heat-inactivated fetal calf serum, the cell number per well was up to 2.1 x 10(5). These monolayer RIN cell cultures were used as a target for the detection of islet cell surface antibodies (ICSA) in the supernatants of hybridomas. The cells were used without fixation to avoid modification of sensitive surface antigens. Poly-D-lysine did not cause non-specific binding of immunoglobulins to the plastic wells as tested with irrelevant monoclonals. The specificity and sensitivity of the method is comparable to indirect immunofluorescence. All mc-ICSA primary screened by indirect immunofluorescence using viable RIN cell suspensions were positive in this CELISA. There was a correlation (r = 0.7; n = 44) between the antibody binding measured by CELISA and the indirect immunofluorescence technique. The advantage of this CELISA is that cell surface structures are well preserved in a viable cell monolayer used as target without chemical fixation. This assay procedure should be generally suitable for the initial screening of monoclonal antibodies to cell surface antigens of cells growing under culture conditions.

Animals↗
PubMed1991

Different multiple reactivity of monoclonal islet cell binding antibodies using indirect immunofluorescence technique on viable cells or cellular ELISA on desiccated cells as target.

Two commonly used methods for screening hybridoma supernatants secreting monoclonal islet cell reactive antibodies (mc-ICRA) were performed to investigate the specificity of the monoclonals established. For this, endothelial, neuroblastoma, murine subcutis and two myeloma cell lines were used as targets in comparison to the insulin-producing rat insulinoma cell line (RIN), either immobilized and permeabilized in cellular enzyme linked immunosorbent assay (CELISA) or in suspension of viable cells in the indirect immunofluorescence test. In addition, rat splenocytes were used for estimating multireactivity of mc-ICRA in ELISA. Using permeabilized target cells, we obtained a high multireactivity of the monoclonal antibodies (mab) tested, indicating a high incidence of molecular mimicry between cytoplasmic antigens of different cell lines. In contrast to CELISA, if only cell surface antigens of viable cells are accessible, detected by the immunofluorescence technique, the high multireactivity is not observed. For investigating the specificity of monoclonals, the complexity of target antigens used must be taken into consideration.

Animals↗
PubMed1990

Cross-infection hazards associated with the use of pumice in dental laboratories.

The bacteriological contamination of pumice slurry in polishing units in a dental clinical area (high risk), a production laboratory (medium risk), and a non-clinical teaching laboratory (low risk), was investigated. Slurry samples taken from all three areas were found to be heavily contaminated with pathogenic organisms. The investigations were repeated following the addition of a disinfectant with both bacteriocidal and virucidal properties to the pumice. Lower bacterial counts were obtained indicating that routine disinfection of pumice slurries is desirable.

Bacteroides↗
PubMed1990

[Binding of monoclonal antibodies to an insulin-producing rat tumor cell (RIN) after partial cell synchronization].

Murine monoclonal antibodies against pancreatic islet cell antigens were generated by somatic cell hybridization and their binding to the insulin-producing rat insulinoma cell line (RIN-5 AH) was evaluated in an indirect immunofluorescence test. An ascites dilution of 1:50 was used for immunostaining using RIN cells harvested from monolayer cultures in RPMI 1640 supplemented with 7.5% fetal calf serum (controls) or test cultures supplemented with 10 mmol/l hydroxyurea to collect the cells in G1/S phase. The five mc-ICSA tested showed a wide variety in their individual binding from 95 up to 30% but their binding was not influenced by hydroxyurea use for synchronization. The antigenic determinant recognized by the anti-glucolipid mc-ICSA 56a F3 was not present on all RIN-5 AH cells. In average, only 35% of the RIN cells showed a striking binding of the monoclonal 56a F3.

Animals↗