PubMed Health⌕ Search

Biomedical subjects

J Wegener

Publications and source records attributed to J Wegener.

At least 19 recordsLinked to original sources
PubMed2007

Analysis of calcium channels by conditional mutagenesis.

Ca2+ influx through various ion channels is an important determinant of the cytosolic Ca2+ concentration, which plays a pivotal role in countless cellular processes. The cardiac L-type Ca2+ channel, Ca(v)1.2, represents a major pathway for Ca2+ entry and is in many cells expressed together with other high- and low-voltage-activated Ca2+ channels. This article will focus on the use of conditional transgenic mouse models to clarify the roles of Ca2+ channels in several biological systems. The phenotypes of conditional Ca2+ channel transgenic mice have provided novel, and often unexpected, insights into the in vivo function of L-type and T-type Ca2+ channels as mediators of signaling between cell membrane and intracellular processes in blood pressure regulation, smooth muscle contractility, insulin secretion, cardiac function, sleep, learning, and memory.

Animals↗
PubMed2006

Bradykinin shifts endothelial fluid passage from para- to transcellular routes.

The signalling peptide bradykinin (BK) is implicated in inflammation and angiogenesis. It promotes fluid transport from blood vessels to interstitial space, and thus facilitates oedema formation. To clarify whether paracellular or transcellular pathways mediate this effect, we investigated the BK-stimulated fluid transport across endothelial monolayers in vitro by comparison of electrical and fluorescence methods. Electrical cell impedance sensing monitored a biphasic response of cell layers to BK with high time resolution: a short decrease (18%, 1 min) was followed by a more sustained increase in paracellular resistance (30%, 10 min). The two phases can be assigned to second messengers of the BK-signalling pathway: Ca(2+) for the decrease and cyclic adenosine monophosphate for the rise of resistance, respectively. Despite tightening of the intercellular clefts, BK increased the fluid permeability by 39%, indicating transcellular fluid transport. Additionally, BK stimulated both in- and outwardly directed membrane trafficking as assessed by vesicular fluid uptake (by 49%) and secretion of von Willebrandt factor (by 85%). In conclusion, the combination of electrical and fluorescence data suggests that BK induces a shift from para- to transcellular fluid transport across endothelium.

Body Fluid Compartments↗
PubMed2004

Critical soft tissue parameters of the zygomatic implant.

AIM: Zygomatic implants have been introduced for the rehabilitation of patients with severe bone defects of the maxilla. The soft tissue aspects of the palatal emergence situation have not been described yet. The aim of this study was to evaluate the incidence and clinical impact of possible periimplant alterations of zygomatic implants. MATERIALS AND METHODS: From 1998 to 2001 all patients with zygomatic implants were included into this study (24 patients, 37 zygomatic implants). One implant was lost in the loading phase giving a survival rate of 97%. Fourteen patients with 20 zygomatic implants fulfilled the inclusion criteria and were all available for the recall examination. Thirteen zygomatic implants were inserted in cases of severe maxillary atrophy, seven in cases of tumour-resection of the maxilla. Clinical examination and microbial analysis using a DNA probe was performed. The implants had a mean time in situ of 598 days (min: 326, max: 914). RESULTS: Colonisation with periodontal pathogens was found at four of the 20 implants. A positive microbiologic result of the periimplant pocket and the maximum pocket probing depth were not statistically related. Nine of the 20 implants showed bleeding on probing, four of these had positive microbiologic results. At sites without bleeding on probing only negative microbiologic samples were found (p=0.026). The mean palatal and mesial probing depth was 1 mm deeper than at the vestibular and distal aspect. Thus at nine out of the 20 implants both, bleeding on probing and pocket probing depth >/=5 mm indicated soft tissue problems resulting in a success rate of only 55%. The patient's history (tumor versus atrophy) or smoking habits seemed not to have influence the situation. CONCLUSION: These soft tissue problems should be taken into account if zygomatic implants are considered as an alternative therapy option in the maxilla.

Alveolar Bone Loss↗
PubMed2002

[Patient satisfaction with dental implants].

BACKGROUND: Almost all implant systems available today achieve success rates of over 90%. The subjective satisfaction of patients with dental implants is only rarely considered in the definition of implant success and there are only limited data on the satisfaction of patients with dental implants in the literature. Therefore, a satisfaction analysis was carried out in a retrospective study. DESIGN OF STUDY: In addition to a clinical investigation, 330 patients with 1,273 implants (Astra, Frialit-2, IMZ TwinPlus and ITI) were asked to answer a standardized questionnaire to rate their personal opinion on the implant success. A total of 199 questionnaires (803 implants, 63%) were returned and could be analyzed. RESULTS: Of the patients, 67% participated in regular follow-up after implantation and 15% indicated postoperative discomfort. Patient satisfaction with the implants (rated on a scale of 1-6) was good or excellent (2 and 1, respectively) in 88% of cases. Satisfaction with the prosthetic treatment was 80%. The whole treatment was considered good or very comfortable by 86% of patients. If necessary, 91% of patients who underwent the procedure would do so again, and 94% would recommend the procedure to someone else. The Spearman correlation analysis did not reveal any statistically significant correlations between the clinical parameters and patient satisfaction scores. DISCUSSION: Despite the high number of patients involved ( n=330), the expected influence of objective clinical parameters on patient satisfaction could not be statistically evaluated. Nevertheless, in addition to the clinical parameters, subjective patient satisfaction should be included in the evaluation of implant success.

Dental Implants↗
PubMed2001

Porcine Choroid plexus epithelial cells in culture: regulation of barrier properties and transport processes.

The epithelial cells of the choroid plexus are the structural basis of the blood-cerebrospinal fluid (CSF)-barrier. Here we summarise our recent efforts to culture those cells mainly on permeable supports in vitro. Isolated from porcine brains, we report a simple protocol for the primary culture using cytosine arabinoside as an additive that is cytotoxic for other cells except the plexus epithelial cells. Enhanced barrier properties are obtained by withdrawal of serum from the culture medium after confluency is reached. Cells improve their polarity, permeability for hydrophilic substrates is lowered, electrical resistance is increased tenfold, and a pH-gradient is built up across the cell monolayer. Polarised secretion of proteins and most importantly fluid secretion into the apical filter compartment was attained and proven to be dependent on the Na(+),K(+)-ATPase activity. Active transport processes (penicillin G, riboflavin, myo-inositol, ascorbic acid) were studied and clearly showed the involvement of the organic anion transporter. The permeability of the barrier was found to be regulated by cyclic adenosine monophosphate (cAMP). Moreover, we report that cell proliferation and differentiation is controlled by components of the extracellular matrix. The present culture model could now be used as an in vitro system to quantify drug transport across the blood-CSF-barrier.

Animals↗
PubMed2001

The quartz crystal microbalance as a novel means to study cell-substrate interactions in situ.

The quartz crystal microbalance (QCM) was first introduced as a mass sensor in gas phase and in vacuum. Since oscillator circuits capable of exciting shear vibrations of quartz resonators under liquid loading have been developed, the QCM became accepted as a new, powerful technique to follow adsorption processes at solid-liquid interfaces in chemical and biological research. Lately, the QCM technique has attracted considerable interest as a novel means to monitor cell-substrate interactions of mammalian cells in vitro. Because the establishment and modulation of cell-substrate contacts is important for many physiological processes, and potent techniques to measure these interactions noninvasively are rare, the present review highlights applications of the QCM technique in this field. The suitability of the QCM device to monitor attachment and spreading of mammalian cells in real time has been well established. The QCM response is dependent on the individual cell type that is examined. In order to identify the sources for these cell-type-specific results of QCM readings, and to understand the information content of the signal, attempts have been made to decompose the overall QCM response into subcellular contributions. The aforementioned subjects, together with a condensed introduction into the QCM technology, are included in this article.

Adsorption↗
PubMed2000

Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces.

This article describes the optimization of an experimental technique referred to as electric cell-substrate impedance sensing (ECIS) to monitor attachment and spreading of mammalian cells quantitatively and in real time. The method is based on measuring changes in AC impedance of small gold-film electrodes deposited on a culture dish and used as growth substrate. Based on experimental data and theoretical considerations we demonstrate that high-frequency capacitance measurements (f = 40 kHz) are most suited to follow the increasing surface coverage of the electrode due to cell spreading. The excellent time resolution of the method allowed an in-depth analysis of cell spreading kinetics under various experimental conditions. Using ECIS we studied the attachment and spreading of epithelial MDCK cells (strain II) on different protein coatings, and investigated the influence of divalent cations on spreading kinetics. We quantified the inhibitory effect of soluble peptides that mimic the recognition sequence of fibronectin and other extracellular matrix proteins (RGDS). We also applied the ECIS technique to monitor the detachment of confluent fibroblastic cell layers (WI38/VA-13) by means of these peptides.

Animals↗
PubMed2000

Barrier function of porcine choroid plexus epithelial cells is modulated by cAMP-dependent pathways in vitro.

In the present paper, we demonstrate that the barrier properties of primary cultured epithelial cells isolated from porcine choroid plexus are regulated by cAMP-dependent signal transduction pathways in vitro. Triggering cAMP-connected cascades in cell layers grown on permeable filters with cAMP-analogues or forskolin led to a significant increase of transepithelial electrical resistances and a pronounced reduction in the permeation rate of a 4 kDa-dextran probe. In dose-response experiments using the cAMP-analogue 8-(4-chlorophenylthio)-cAMP transepithelial electrical resistances were observed to increase above a threshold concentration ranging between 10(-5.5) and 10(-5) M. Additional impedance studies performed with confluent cell layers grown on gold-film electrodes revealed that the observed changes in transepithelial resistances and presumably also in macromolecular permeation rates were not entirely caused by a reinforcement of intercellular junctions but also contained contributions from changes in the cell-substrate adhesion pattern. These inherent contributions to the electrical resistance and macromolecular permeability are caused by a restricted diffusion pathway between basal plasma membrane and culture substrate that have to be considered in data analysis, especially when leaky cell layers on filter substrates with low pore densities are used.

Animals↗
PubMed2000

Analysis of the composite response of shear wave resonators to the attachment of mammalian cells.

The suitability of the quartz crystal microbalance (QCM) technique for monitoring the attachment and spreading of mammalian cells has recently been established. Different cell species were shown to generate an individual response of the QCM when they make contact with the resonator surface. Little is known, however, about the underlying mechanisms that determine the QCM signal for a particular cell type. Here we describe our results for different experimental approaches designed to probe the particular contributions of various subcellular compartments to the overall QCM signal. Using AC impedance analysis in a frequency range that closely embraces the resonators' fundamental frequency, we have explored the signal contribution of the extracellular matrix, the actin cytoskeleton, the medium that overlays the cell layer, as well as the liquid compartment that is known to exist between the basal plasma membrane and the culture substrate. Results indicate that the QCM technique is only sensitive to those parts of the cellular body that are involved in cell substrate adhesion and are therefore close to the resonator surface. Because of its noninvasive nature, sensitivity, and time resolution, the QCM is a powerful means of quantitatively studying various aspects of cell-substrate interactions.

3T3 Cells↗
PubMed1999

Analysis and occurrence of estrogenic hormones and their glucuronides in surface water and waste water in The Netherlands.

An analytical procedure was developed that enables routine analysis of four estrogenic hormones in concentrations below 1 ng/l in surface water and waste water. The recovery was 88-98% with a limit of detection of 0.1-2.4 ng/l depending on the compound and the matrix measured. This method was used to determine the occurrence of 17 beta-estradiol, 17 alpha-estradiol, estrone and 17 alpha-ethinylestradiol in the aquatic environment in The Netherlands. The data show that estrogenic hormones can be detected at low concentrations (up to 6 ng/l) at some locations in surface water. In selected effluents of waste water treatment plants estrone and 17 beta-estradiol were detected in concentrations in the ng/l range. Concentrations of 17 alpha-estradiol and the contraceptive 17 alpha-ethinylestradiol were in most of these samples below the limit of detection. Hormone glucuronides were not detected in most surface water and effluents.

Animals↗
PubMed1999

[Prognosis and prognostic factors of endosseous implants in the irradiated jaw].

In comparison to tumor patients not receiving radiotherapy, the rehabilitation of masticatory function after head and neck irradiation is limited due to radiation-induced caries, radioxerostomia, and the risk of osteoradionecrosis. This study focused on implants in the irradiated jaw and on the evaluation of the prognosis and the effect of potential factors on the prognosis. The retrospective study covered 197 implants (47 patients) from 1988 to 1997. The implant prognosis was determined by implant survival statistics (Kaplan-Meier). Losses not related to the implants were censored. In addition, groups were formed according to factors potentially affecting the prognosis. The significance of differences in the groups relative to survival were tested using the log-rank test. Twelve (6.1%) implants from a total of 197 were lost due to peri-implantitis, and eight (4.1%) due to possible biomechanical stress. A total of 52 losses (26.4%) due to death of patients and two (1.0%) due to resection of the jaw were censored; 111 (56.3%) implants remained at recall and the average interval was 33 months. The rates of implant survival (Kaplan-Meier) after 1 and 2 years were 95%, after 3 and 4 years 92%, and after 5 and 6 years 72%. The univariate analysis of group comparisons showed a significantly lower rate of loss after perimplant flap reconstruction (p = 0.036). There was no effect due to the doses of irradiation (p = 0.16), chemotherapy (p = 0.90), or peri-implant osteoplasty (p = 0.84). Although none of the implants inserted before radiotherapy had to be explanted, the implant survival difference in the very heterogeneous groups was not significant (preirradiation, n = 29; postirradiation: n = 156; p = 0.13). According to the literature, the rate of survival of teeth which were sound before radiotherapy (1 year, 75%; 5 years, 45%) was distinctly lower than the survival of enossal implants (1 year, 95%; 5 years, 72%). The high-quality rehabilitation of masticatory function with implant-based protheses is the preferred method of treatment for irradiated tumor patients. In addition, contraindications for enossal implants were ruled out for all studied factors affecting prognosis.

Adult↗
PubMed1999

[The maxilla--a poor implant site?].

In the maxilla, it is sometimes necessary to use implants to achieve a good prosthetic result. However, an increased failure rate of maxillary implants seems to be a common clinical experience. This experience was investigated in a retrospective statistical analysis. In a retrospective study of 665 patients between 1987 and 1997, 2484 implants were examined. The implants in the upper jaw were compared to those in the lower jaw. Implants with and without autogenous bone grafts and fixtures in patients with alveolar ridge atrophy, tumor, or trauma were explored. In particular, the data obtained from the orthopantomograms after completion of the prosthetic superstructures were controlled for peri-implantary bone loss evaluation. A total of 40% of the fixtures were placed in the maxilla, and 30% of these implants were combined with a local or an iliac bone graft. Out of 2484 implants, 207 were lost, largely during the healing period. The failure rates in the groups of patients with alveolar ridge atrophy, tumor, and trauma were between 6 and 27.5%, and the differences between these groups were higher than the difference between the upper and the lower jaw. Especially in the maxilla and after osteoplasty, horizontal peri-implantary bone loss was increased, with some cases of dramatic bone loss and decrease of the osseous integration of the fixtures. There was no statistic evidence of a significantly higher failure rate in the upper jaw than in the lower jaw. However, besides the rate of implant loss, additional markers should also be considered for adequate evaluation of implant prognosis in the maxilla.

Alveolar Bone Loss↗
PubMed1999

Cell adhesion monitoring using a quartz crystal microbalance: comparative analysis of different mammalian cell lines.

The quartz crystal microbalance (QCM) has been widely accepted as a sensitive technique to follow adsorption processes in gas as well as in liquid environments. However, there are only a few reports about the use of this technique to monitor the attachment and spreading of mammalian cells onto a solid support in culture. Using a QCM-setup we investigated the time course of cell attachment and spreading as a function of seeding density for three widespread and frequently used cell lines (MDCK strains I and II and Swiss 3T3-fibroblasts). Results were found to be in good agreement with the geometrical properties of the individual cell types. The shifts of the resonance frequency associated with confluent cell layers on top of the quartz resonators were found to be dependent on the cell species [MDCK-I: (320 +/- 20) Hz; MDCK-II: (530 +/- 25) Hz; 3T3: (240 +/- 15) Hz] reflecting their individual influence on the shear oscillation of the resonator. These findings are discussed with respect to the basic models of materials in contact with an oscillating quartz resonator. We furthermore showed by inhibition-assays using soluble RGD-related peptides, that only specific, integrin mediated cell adhesion is detected using this QCM approach, whereas the sole presence of the cellular body in close vicinity to the resonator surface is barely detectable.

Animals↗
PubMed1999

Use of electrochemical impedance measurements to monitor beta-adrenergic stimulation of bovine aortic endothelial cells.

Due to the high permeability of endothelial cell layers derived from macrovascular vessels, precise determination of their barrier function towards ion movement requires refined experimental techniques. We thus cultured bovine aortic endothelial cells (BAEC) directly on thin gold-film electrodes and measured the electrochemical impedance to study their passive electrical properties in general and during beta-adrenergic stimulation. Impedance spectra (10-2.10(6) Hz) of confluent cell monolayers revealed that the electrical characteristics of the cells can be modelled by a simple resistor-capacitor parallel network. Under control conditions the overall resistance of confluent BAEC monolayers was 3.6+/-0.6 Omega.cm2 (n=30) and the capacitance was 0. 6+/-0.1 microF/cm2. Both quantities are discussed with respect to morphological characteristics of these cells. Stimulation of BAECs with the synthetic beta-adrenoceptor agonist isoproterenol leads to a concentration-dependent, highly specific increase of the cell layer resistance characterized by a concentration for half-maximal response (EC50) of 0.3+/-0.1 microM. The cell layer capacitance, however, remained unaffected. Using impedance measurements at a single frequency, we analysed the response of BAECs to treatment with isoproterenol in comparison with several chemically unrelated compounds known to stimulate the adenosine 3',5'-cyclic monophosphate (cAMP)-dependent signal transduction cascade. These studies confirmed that the enhancement of the cell layer resistance after beta-adrenergic stimulation is mediated by an increase in intracellular cAMP.

Adenylyl Cyclases↗
PubMed1999

Acting without asking: an ethical analysis of the Food and Drug Administration waiver of informed consent for emergency research.

This article summarizes the current requirements for a waiver of informed consent for emergency research and analyzes ethical issues that are involved. Researchers who intend to apply for a waiver of informed consent for emergency research must understand that they are asking for the major protector of human subjects, the informed consent process, to be removed. In its absence, other protections are required. These include communication with the community, family members, and others close to the patient. It is unlikely that even these additional protections can equal that afforded by an adequate consent process. With this understanding, investigators' actions must be in accordance with the highest ethical standards. It is therefore necessary for investigators to know the ethical implications of conducting controlled research without the subjects' consent.

Community-Institutional Relations↗
PubMed1998

Hydrocortisone reinforces the blood-brain properties in a serum free cell culture system.

The increasing number of newly developed drugs demands for functional in vitro models of the blood-brain barrier to determine their brain uptake. Cultured cerebral capillary endothelial cells are considered to be such a model, however in serum containing media they exhibit low electrical resistances and high permeabilities compared to the in vivo situation. Here we report the establishment of a serum-free cell culture model. Withdrawal of serum already caused a twofold increase of transendothelial resistance (TER), which in presence of serum is about 100-150 Omega x cm2. We tested several supplements and found that hydrocortisone is a potent stimulator for the formation of barrier properties. TERs up to 1000 Omega x cm2 were measured in the presence of physiological relevant hydrocortisone concentrations. In correspondence to the TER increase hydrocortisone decreased cell monolayer permeability for sucrose down to 5x10(-7) cm/s, which is close to the in vivo value of 1.2x10(-7) cm/s and by a factor of five lower compared to cultures without hydrocortisone and in presence of serum.

Animals↗
PubMed1998

Hydrocortisone reinforces the blood-brain barrier properties in a serum free cell culture system.

The increasing number of newly developed drugs demands for functional in vitro models of the blood-brain barrier to determine their brain uptake. Cultured cerebral capillary endothelial cells are considered to be such a model, however in serum containing media they exhibit low electrical resistances and high permeabilities compared to the in vivo situation. Here we report the establishment of a serum-free cell culture model. Withdrawal of serum already caused a twofold increase of transendothelial resistance (TER), which in presence of serum is about 100-150 omega.cm2. We tested several supplements and found that hydrocortisone is a potent stimulator for the formation of barrier properties. TERs up to 1000 omega.cm2 were measured in the presence of physiological relevant hydrocortisone concentrations. In correspondence to the TER increase hydrocortisone decreased cell monolayer permeability for sucrose down to 5 x 10(-7) cm/s, which is close to the in vivo value of 1.2 x 10(-7) cm/s and by a factor of five lower compared to cultures without hydrocortisone and in presence of serum.

Animals↗