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U Klein

Publications and source records attributed to U Klein.

At least 19 recordsLinked to original sources
PubMed2001

Heterologous inhibition of G protein-coupled receptor endocytosis mediated by receptor-specific trafficking of beta-arrestins.

We have observed an unexpected type of nonreciprocal "cross-regulation" of the agonist-induced endocytosis of G protein-coupled receptors by clathrin-coated pits. Isoproterenol-dependent internalization of beta2-adrenergic receptors in stably transfected HEK293 cells was specifically blocked (>65% inhibition) by vasopressin-induced activation of V2 vasopressin receptors co-expressed at similar levels. In contrast, activation of beta2 receptors caused no detectable effect on V2 receptor internalization in the same cells. Several pieces of evidence suggest that this nonreciprocal inhibition of endocytosis is mediated by receptor-specific intracellular trafficking of beta-arrestins. First, previous studies showed that the activation of V2 but not beta2 receptors caused pronounced recruitment of beta-arrestins to endocytic membranes (Oakley, R. H., Laporte, S. A., Holt, J. A., Barak, L. S., and Caron, M. G. (1999) J. Biol. Chem. 274, 32248-32257). Second, overexpression of arrestin 2 or 3 (beta-arrestin 1 or 2) abolished the V2 receptor-mediated inhibition of beta2 receptor internalization. Third, mutations of the V2 receptor that block endomembrane recruitment of beta-arrestins eliminated the V2 receptor-dependent blockade of beta2 receptor internalization. These results identify a novel type of heterologous regulation of G protein-coupled receptors, define a new functional role of receptor-specific intracellular trafficking of beta-arrestins, and suggest an experimental method to rapidly modulate the functional activity of beta-arrestins in intact cells.

Arginine Vasopressin↗
PubMed2001

May pentoxifylline improve lung function after one-lung flooding?

One-lung flooding makes the intraoperative sonography of round pulmonary lesions possible. During the flooded phase, the flooded lung suffers a significant reduction in perfusion. After ischemia and reperfusion, neutrophil granulocytes lead to further tissue injuries. A study was made on four animals to determine whether administration of pentoxifylline--a potent inhibitor of granulocyte adhesion to the endothelium--improves lung function after one-lung flooding. Two animals were subjected to thoracotomy with extended hemodynamic monitoring. Thoracoscopy was performed on two other animals, which were extubated after the flooding liquid was drained and survived for 24 degrees h. A bolus of 1 mg/kg of pentoxifylline was administered at the time of thoracotomy/thoracoscopy. followed by continuous infusion of pentoxifylline at a rate of 1.5 mg/kg per hour until 30 degrees min after reventilation (thoracotomy), or until extubation, respectively. The control group consisted of animals employed in previous experiments. Except for pentoxifylline administration, they were subjected to identical experimental conditions. The control group for the thoracotomy experiment comprised 14 animals, that for the thoracoscopy experiment three animals. The experiments proved that 30 degrees min after the draining of the flooding liquid and reventilation, all four pentoxifylline-treated animals had a higher partial arterial oxygen pressure and a lower pulmonary shunt volume compared with the control animals. In the two animals that survived, a positive effect on lung function was no longer detectable 24 degrees h after extubation. The administration did not lead to a drop in the pulmonary arterial pressure and did not cause any hemodynamic changes other than a moderate tachycardia.

Animals↗
PubMed2001

[Urapidil for treatment of postanesthetic shivering following general anesthesia. A placebo controlled pilot study].

OBJECTIVE: Postanaesthetic shivering is common during recovery from general anaesthesia. Therefore we studied whether urapidil suppresses postanaesthetic shivering. METHODS: With written informed consent and approval of the local ethics committee, patients (ASA I-II) recovering from general anaesthesia were monitored for 1 h. Patients with continuous shivering for a period of 5 min were randomly treated either with 5 ml placebo (isotonic saline) or 25 mg urapidil in a double-blind trial. This treatment procedure was repeated if shivering did not stop. A complete suppression of shivering was appraised as a sufficient treatment. RESULTS: Shivering occurred in 20 of the patients studied and urapidil stopped shivering in 7 out of the 10 treated patients, whereas the placebo stopped shivering in only 2 out of 10 patients (P < 0.05). CONCLUSION: In a placebo controlled trial, it was demonstrated that postanaesthetic shivering can be successfully treated by urapidil in 70% of the patients.

Adrenergic alpha-Antagonists↗
PubMed2001

Specific sequence elements in the 5' untranslated regions of rbcL and atpB gene mRNas stabilize transcripts in the chloroplast of Chlamydomonas reinhardtii.

Using a series of point mutations in chimeric reporter gene constructs consisting of the 5' regions of the Chlamydomonas chloroplast rbcL or atpB genes fused 5' to the coding sequence of the bacterial uidA (GUS) gene, RNA-stabilizing sequence elements were identified in vivo in the 5' untranslated regions (5' UTRs) of transcripts of the chloroplast genes rbcL and atpB in Chlamydomonas reinhardtii. In chimeric rbcL 5' UTR:GUS transcripts, replacement of single nucleotides in the 10-nt sequence 5'-AUUUCCGGAC-3', extending from positions +38 to +47 relative to the transcripts' 5' terminus, shortened transcript longevity and led to a reduction in transcript abundance of more than 95%. A similar mutational analysis of atpB 5' UTR:GUS transcripts showed that the 12-nt atpB 5' UTR sequence 5'-AUAAGCGUUAGU-3', extending from position +31 to position +42, is important for transcript stability and transcript accumulation in the chloroplast of Chlamydomonas. We discuss how the 5' UTR sequence elements, which are predicted to be part of RNA secondary structures, might function in RNA stabilization.

5' Untranslated Regions↗
PubMed2001

Oxygenation during one-lung ventilation: the effects of inhaled nitric oxide and increasing levels of inspired fraction of oxygen.

UNLABELLED: We studied whether inhaled nitric oxide (NO) would improve arterial oxygen tension (PaO(2)) and reduce the occurrence of oxygen saturation of hemoglobin (O(2)Hb) < 90% during one-lung ventilation (OLV). One-hundred-fifty-two patients were ventilated either with or without NO (20 ppm) with an inspired fraction of oxygen (FIO(2)) of either 0.3, 0.5, or 1.0 during OLV. Anesthesia was induced and maintained with propofol, remifentanil, and rocuronium IV, and lung separation was achieved with a double-lumen tube. During OLV, we set positive end-expiratory pressure at 5 cm H(2)O, peak pressure at 30 cm H(2)O, and end-tidal CO(2) at 30 mm Hg. The nonventilated lung was opened to room air and collapsed. During OLV, three consecutive measurements were performed every 10 min. The operated lung was temporarily ventilated if pulse oximetric saturation (SpO(2)) decreased to < 91%. SpO(2) <9 1% occurred in 2 of the 152 patients. SpO(2) overestimated O(2)Hb by 2.9% +/- 0.1%. NO failed to improve oxygenation or alter occurrence of O(2)Hb < 90% during OLV across all time points and all levels of FIO(2). Increasing FIO(2) increased oxygenation and decreased occurrence of O(2)Hb < 90% (P: < 0.001). At FIO(2) = 1, PaO(2) was higher (P < 0.01) and O(2)Hb < 90% rate tended to be lower (P = 0.1) during right versus left lung ventilation. PaO(2) was higher in patients undergoing pneumonectomy and lobectomy than in those undergoing metastasectomy or video-assisted operations (P < 0.05). IMPLICATIONS: Inhaled nitric oxide failed to improve oxygenation during one-lung ventilation. Oxygenation during one-lung ventilation was improved with increasing levels of FIO(2) during ventilation of the right versus the left lung and with increasing pathology of the nonventilated lung.

Adult↗
PubMed2000

Disruption of the Rev3l-encoded catalytic subunit of polymerase zeta in mice results in early embryonic lethality.

Polymerase zeta (Pol zeta) is an error-prone DNA polymerase [1], which in yeast is involved in trans-lesion synthesis (TLS) and is responsible for most of the ultraviolet (UV) radiation-induced and spontaneous mutagenesis [2-4]. Pol zeta consists of three subunits: REV1, a deoxycytidyl-transferase [5]; REV7, of unclear function [6]; and REV3, the catalytic subunit. REV3 alone is sufficient to carry out TLS, but association with REV1 and REV7 enhances its activity [5, 7]. Experiments using human cells treated with UV radiation indicate also that mammalian Pol zeta is involved in TLS [7]. The peculiar mutagenic activity of Pol zeta [4,7,8] suggests a possible role in somatic hypermutation of immunoglobulin (Ig) genes [9]. Here, we report that, unlike in yeast where the REV3 gene is not essential for life [4], disruption of the mouse homologue (Rev3l) resulted in early embryonic lethality. In Rev3l(-/-) embryos, no haematopoietic cells other than erythrocytes could be identified in the yolk sac. Rev3l(-/-) haematopoietic precursors were unable to expand in vitro and no haematopoietic cells could be derived from the intraembryonic haematogenic compartment (splanchnopleura). Fibroblasts could not be derived from the Rev3l(-/-) embryos, and Rev3l(-/-) embryonic stem (ES) cells could not be obtained. This is the first evidence that an enzyme involved in TLS is critical for mammalian development.

Animals↗
PubMed2000

Mice reconstituted with DNA polymerase beta-deficient fetal liver cells are able to mount a T cell-dependent immune response and mutate their Ig genes normally.

The ubiquitously expressed, error-prone DNA polymerase beta (polbeta) plays a role in base excision repair, and the involvement of this molecule in the nonhomologous end joining (NHEJ) process of DNA repair has recently been demonstrated in yeast. Polbeta-deficient mice are not viable, and studies on conditional mutants revealed a competitive disadvantage of polbeta(-/-) vs. wild-type cells. We show here that polbeta-deficient mice survive up to day 18.5 postcoitum, but die perinatally; a circumstance that allowed the investigation of a potential role of polbeta in lymphocyte development by transfer of fetal liver cells (FLC) derived from polbeta(-/-) embryos into lethally irradiated hosts. FLC transfers using mutant cells lead to an almost normal reconstitution of the lymphocyte compartment, indicating that polbeta-deficiency does not prevent V(D)J recombination, which is known to employ factors of the NHEJ pathway. Mice reconstituted with polbeta(-/-) FLC mount a normal T cell-dependent immune response against the hapten (4-hydroxy-3-nitrophenyl) acetyl (NP). Moreover, germinal center B cells from NP-immunized reconstituted mice show normal levels and patterns of somatic point mutations in their rearranged antibody genes, demonstrating that polbeta is not critically involved in somatic hypermutation.

Animals↗
PubMed2000

CD5-positive B cells in healthy elderly humans are a polyclonal B cell population.

B cell chronic lymphocytic leukemia (B-CLL) is a disease of the elderly and is characterized by a malignant clone of CD5+ B cells. In old mice, clonal expansions of CD5+ B cells are a common feature, and these animals frequently develop B-CLL. To investigate whether clonal expansion of CD5+ B cells also occurs in elderly humans, predisposing for the development of B-CLL, we analyzed VH gene rearrangements of CD5+ B cells from blood samples of four healthy, 65-82-years-old volunteers as markers of clonality. CD5+ and CD5-B cells were obtained by cell sorting, CDRIII of rearranged VH genes were amplified by polymerase chain reaction, and fragment length analysis was performed. All samples demonstrated a polyclonal pattern of VH gene length distribution. In addition, VH gene rearrangements were amplified and sequenced from sorted single cells of two of the donors. No clonally related CD5+ or CD5- B cells were observed. Thus, development of dominant clones of CD5+ peripheral blood B cells is unlikely to be a common trait of elderly individuals.

Aging↗
PubMed2000

One-lung flooding for video-assisted thoracoscopic surgery in animal experiments on pigs--oxygenation and intrapulmonary shunt.

Unilateral flooding of the lung after intubation with a double-lumen tube makes intraoperative sonography of the lung during video-assisted thoracoscopic surgery possible. After flooding with 15 ml/kg, the arterial partial oxygen pressure (with FiO2=1.0) is higher than that in total atelectasis by about 100 mmHg, while it is only slightly less than that during bilateral lung ventilation. Compared to total atelectasis, lung flooding reduces the pulmonary right-to-left shunt volume. The pulmonary function normalizes within 8 h after the operation.

Air↗
PubMed2000

Comparison of cisatracurium-induced neuromuscular block at the masseter and adductor pollicis muscle.

Adequate relaxation of the masseter muscle is important during endotracheal intubation and for the patency of a patient's airway during recovery from anaesthesia. We evaluated onset and recovery from cisatracurium-induced neuromuscular block at the masseter and adductor pollicis muscles. Thirty patients were randomly allocated to receive either 0.1 or 0.15 mg kg(-1) cisatracurium. The evoked response was measured at both muscles using acceleromyography. Onset time was significantly shorter at the masseter muscle than at the adductor pollicis (0.1 mg kg(-1) cisatracurium: 155+/-52 vs. 229+/-44 s; 0.15 mg kg(-1) cisatracurium: 105+/-24 vs. 174+/-35 s). Following 0.1 mg kg(-1) cisatracurium, recovery to a TOF-ratio of 0.7 was faster at the masseter compared to the adductor pollicis (P < 0.05). In the 0.15 mg kg(-1) cisatracurium group recovery of T1 to 75% of control and to a TOF-ratio of 0.7 occurred sooner at the masseter (P < 0.05). We conclude that onset and recovery from cisatracurium neuromuscular block occurs more rapidly at the masseter than at the adductor pollicis. It appears unlikely that residual paralysis is present at the masseter once neuromuscular function at the adductor pollicis has completely recovered.

Adult↗
PubMed2000

Wet-to-dry ratio of lung tissue and surfactant outwash after one-lung flooding.

Unilateral flooding of the lung after intubation with a double-lumen tube enables intraoperative sonography of the lung tissue. The flooding restricts the pulmonary blood stream of the flooded lung in a relevant degree and thus reduces the right-to-left shunt volume. The deficient perfusion of the lung tissue during the flooded phase might cause capillary permeability disorders and secondary oedema development. This can be determined by examination of extravascular lung water (EVLW) after draining and reventilation of the flooded lung. Although one-time unilateral lung flooding must be distinguished from bronchopulmonary lavage, it is interesting to study the effects of flooding on the surfactant system. The wet-to-dry ratio of the lung tissues of 13 female pigs was ascertained at different times following one-lung flooding (1 to 11 weeks). A trend towards an increased wet-to-dry ratio in the previously flooded lung was found only in the tissue samples taken 1 h after reventilation. After only 24 h, the two lungs no longer differed in their wet-to-dry ratio. In six pigs, the phospholipid content of the drained flooding liquid was determined. It was shown that the surfactant loss caused by flooding was maximally 47% of the calculated surfactant pool of the respective lung.

Animals↗
PubMed1999

Phenotypic and molecular characterization of human peripheral blood B-cell subsets with special reference to N-region addition and J kappa-usage in V kappa J kappa-joints and kappa/lambda-ratios in naive versus memory B-cell subsets to identify traces of receptor editing processes.

We identified a population of IgM+IgD+ B-cells in the peripheral blood (PB) of humans that express somatically mutated V-region genes like classical class switched or IgM-only memory B-cells and comprise around 15% of PB B-cells in adults. Mutated IgM+IgD+ cells differ from unmutated naive IgM+IgD+ cells in that they express the CD27 cell surface antigen. In addition, a very small subset of IgD-only B-cells was identified in the PB that carried rearranged VH-genes with an extremely high load of somatic mutations (up to 60 mutations per gene). A common characteristic of the four somatically mutated subsets, which altogether comprise 40% of PB B-lymphocytes in adults, is the surface expression of CD27. This antigen may thus represent a general marker for memory B-cells in the human. Somatically mutated and unmutated PB B-cell subsets were analyzed for N-region addition and J kappa-usage in V kappa J kappa-joints, and in addition for the respective kappa/lambda-ratios: N-nucleotides could be identified in a large fraction of V kappa-regions of all B-cell subsets, indicating that N-region insertion already occurs in the pre-germinal center (GC) phase of B-cell development. Both the J kappa-usage in expressed V kappa J kappa-joints and the kappa/lambda-ratio from somatically mutated B-cells do not differ substantially from those of the unmutated cells, so that in terms of these parameters, a contribution of secondary V kappa J kappa-rearrangements in shaping the memory B-cell repertoire is not detectable.

Adult↗
PubMed1999

Onset of neuromuscular block at the masseter and adductor pollicis muscles following rocuronium or succinylcholine.

PURPOSE: To compare the onset time of two different rocuronium doses (0.6 and 0.9 mg x kg(-1)) and succinylcholine (1.5 mg x kg(-1), preceeded by 0.06 mg x kg(-1) rocuronium) at the masseter and the adductor pollicis muscle. METHODS: In a randomized study, 60 ASA I or II adult women, 18-65 yr of age, were anesthetized with propofol and fentanyl and nitrous oxide in oxygen. Neuromuscular monitoring was performed using acceleromyography simultaneously on the masseter and adductor pollicis. Onset time was measured at both muscles using supramaximal 0.1 Hz single twitch stimulation (square-wave pulse 0.2 msec duration). RESULTS: In all patients, complete neuromuscular block occurred at the masseter and adductor pollicis muscles. Lag-time and onset time were faster at the masseter that at the adductor pollicis muscle in both rocuronium-groups (P < 0.01) and in the succinylcholine-group (P < 0.01). Furthermore, onset time was more rapid after 0.9 mg x kg(-1) rocuronium (65 +/- 7 s) than after succinylcholine (83 +/- 19 sec) at the AP (P < 0.05), but did not differ at the masseter (33 +/- 6 vs 36 +/- 7 sec). CONCLUSIONS: Following rocuronium and succinylcholine, onset time is faster at the masseter than at the adductor pollicis muscle.

Adolescent↗
PubMed1999

[Cisatricurium in the orbicularis oculi muscle. Comparisn of the neuromuscular action of cisatracurium and atracurium in the orbicularis oculi muscle and the adductor pollicis muscle].

OBJECTIVES: Muscle relaxants have different pharmacodynamic profiles in various muscles. Therefore, results obtained for one muscle cannot be extrapolated to other muscles. In the adductor pollicis muscle cisatracurium exerts a pharmacodynamic profile comparable to atracurium, despite the known difference in onset time. However, studies evaluating the neuromuscular effect of cisatracurium in different muscles are lacking. Accordingly, this study compares the pharmacodynamic profile of cisatracurium and atracurium in the orbicularis oculi muscle (OO) - which shows a neuromuscular course similar to the diaphragm and the laryngeal muscles - and the adductor pollicis muscle (AP). METHODS: Forty-five patients (ASA I-II), scheduled for elective spinal surgery were anaesthetized with propofol and fentanyl. Endotracheal intubation was performed without using a muscle relaxant. Neuromuscular transmission was monitored using acceleromyography in both muscles. Patients received 0.1 mg/kg (2x ED(95)) or 0.15 mg/kg (3x ED(95)) cisatracurium, or 0.5 mg/kg atracurium (2x ED(95)) at random. Onset and recovery times were measured according to the recommendation of the Copenhagen Consensus Conference. RESULTS: Onset time was significantly shorter in the OO than in the AP following 0.15 mg/kg cisatracurium and 0.5 mg/kg atracurium (P<0.05). No differences in onset time between the two muscles were found after 0.1 mg/kg cisatracurium. The recovery of T(1) to 10% of its control was completed sooner in the OO than in the AP in all three groups (P<0.05). CONCLUSIONS: Cisatracurium shows a dose-dependent shorter onset time in the OO than in the AP. This is consistent with the current view that the onset of non-depolarizing neuromuscular blockers is more rapid in the OO than in the AP. However, at least a dose of 3x ED(95) of cisatracurium was necessary to show a difference in onset time between both muscles. In contrast, atracurium is reported to lead to a significantly shorter onset of neuromuscular block in the OO following 2x the ED(95). The more rapid recovery of T(1) to 10% of its control in all three groups in the OO is due to the relative resistance of this muscle to muscle relaxants.

Adult↗