Biomedical subjects

T Winkler

Publications and source records attributed to T Winkler.

At least 19 recordsLinked to original sources

PubMed2003

Modeling kinetics of infused 13NN-saline in acute lung injury.

A mathematical model was developed to estimate right-to-left shunt (Fs) and the volume of distribution of 13NN in alveolar gas (VA) and shunt tissue (Vs). The data obtained from this model are complementary to, and obtained simultaneously with, pulmonary functional positron emission tomography (PET). The model describes 13NN kinetics in four compartments: central mixing volume, gas-exchanging lung, shunting compartment, and systemic recirculation. To validate the model, five normal prone (NP) and six surfactant-depleted sheep in the supine (LS) and prone (LP) positions were studied under general anesthesia. A central venous bolus of 13NN-labeled saline was injected at the onset of apnea as PET imaging and arterial 13NN sampling were initiated. The model fit the tracer kinetics well (mean r2 = 0.93). Monte Carlo simulations showed that parameters could be accurately identified in the presence of expected experimental noise. Fs derived from the model correlated well with shunt estimates derived from O2 blood concentrations and from PET images. Fs was higher for LS (54 +/- 18%) than for LP (5 +/- 4%) and NP (1 +/- 1%, P < 0.01). VA, as a fraction of PET-measured lung gas volume, was lower for LS (0.18 +/- 0.09) than for LP (0.96 +/- 0.28, P < 0.01), whereas Vs, as a fraction of PET-measured lung tissue volume, was higher for LS (0.46 +/- 0.26) than for LP (0.05 +/- 0.08, P < 0.01). The main conclusions are as follows: 1) the model accurately describes measured arterial 13NN kinetics and provides estimates of Fs, and 2) in this animal model of acute lung injury, the fraction of available gas volume participating in gas exchange is reduced in the supine position.

Algorithms↗

PubMed2003

[A case of symptomatic paroxysmal kinesigenic dsykinesia with primary central nervous system lymphoma].

Paroxysmal dyskinesias are rare movement disorders. In one fourth of cases, a symptomatic cause can be found, e.g., brain ischemia or inflammatory diseases. We report a unique case of secondary paroxysmal kinesigenic dyskinesia (PKD). The 37-year-old patient suffered from a primary CNS lymphoma invading the right thalamus. Consecutively, he showed dystonic-athetoid movements in the left half of the body. The involuntary movements had a duration of approximately 20 s and occurred more than 100 times per day. Carbamazepine treatment showed excellent results. This is the first reported case of a symptomatic paroxysmal dyskinesia induced by a primary CNS lymphoma.

Adult↗

PubMed2002

Topical application of dynorphin A (1-17) antiserum attenuates trauma induced alterations in spinal cord evoked potentials, microvascular permeability disturbances, edema formation and cell injury: an experimental study in the rat using electrophysiological and morphological approaches.

Dynorphin is a neuropeptide that is present in high quantities in the dorsal horn of the spinal cord. The peptide is actively involved in pain processing pathways. However, its involvement in spinal cord injury is not well known. Alteration in dynorphin immunoreactivity occurs following a focal trauma to the rat spinal cord. Infusion of dynorphin into the intrathecal space of the cord results in ischemia, cell damage and abnormal motor function. Antibodies to dynorphin when injected into the intrathecal space of the spinal cord following trauma improve motor recovery, reduce edema and cell changes. However, influence of dynorphin on trauma induced alteration in spinal cord bioelectrical activity is still not known. Spinal cord evoked potentials (SCEP) are good indicator of spinal cord pathology following trauma. Therefore, in present investigation, influence of dynorphin antibodies on trauma induced changes in SCEP were examined in our rat model. In addition, spinal cord edema formation, microvascular permeability disturbances and cell injury were also investigated. Our results show that topical application of dynorphin antiserum (1 : 200) two min before injury markedly attenuated the SCEP changes immediately after injury. In the antiserum treated animals, a significant reduction in the microvascular permeability, edema formation and cell injury was observed in the traumatised spinal cord. These observations suggest that (i). dynorphin is involved in the altered bioelectrical activity of the spinal cord following trauma, (ii). the peptide actively participates in the pathophysiological processes of cell injury in the spinal cord trauma, and (iii). the dynorphin antiserum has potential therapeutic value for the treatment of spinal cord injuries.

Animals↗

PubMed2002

A pTalpha-negative subpopulation of CD25+ TN thymocytes revealed by a transgenic marker.

We have recently generated 5'lambda5-huTAC mice, which express the human CD25 (huTAC) gene under the control of the 5'-flanking region of the mouse lambda5-gene. The huTAC-transgene was expressed in pre-B cells but neither in mature B cells nor in T cells of these mice. In this report we demonstrate that the transgene is also transiently expressed by adult CD25+ CD3-CD4-CD8- (triple negative, TN) thymocytes and in fetal thymocytes. The huTAC+, in contrast to the huTAC- subpopulation of the CD44+CD25+ TN cells, was unexpectedly found not to express the pTalpha-gene. Still the huTAC+CD44+CD25+ TN cells reconstituted the development of both alphabeta and gammadelta lineage cells equally efficiently as the pTalpha-expressing huTAC- fraction, demonstrating that this pTalpha-negative subpopulation contained precursors for both T-cell lineages. Single cell reverse transcription-polymerase chain reaction (RT-PCR) experiments demonstrated that also in normal mice only a fraction of CD44+CD25+ and CD44-CD25+ TN cells expressed this gene. Taken together, these data indicate that huTAC transgene expression revealed a truly pTalpha-negative fraction of the CD44+CD25+ TN cells. The observation that not all precursors in the CD25+ TN population express the pTalpha-gene has important implications for the understanding of early T-cell development and T-cell lineage commitment.

Animals↗

PubMed2002

Antigenic targets--workshop report.

Several excellent animal models for systemic lupus erythematosus are available to study the pathogenetic mechanisms of anti-DNA antibodies, immunological mechanisms for their origin and the influence of susceptibility genes for the development of anti-DNA antibodies. Furthermore several transgenic mice have been generated to study B cell tolerance against DNA. During the meeting recent progress on animal models for systemic lupus erythematosus was discussed.

Animals↗

PubMed2001

Towards a general procedure for sequencing single DNA molecules.

In this paper we report on the latest technical advances towards single molecule sequencing, a useful method currently developed especially for fast and easy de novo sequencing. Different approaches for complete labeling of DNA with fluorescent dyes are described. In addition, the experimental set-up for the sequencing process is shown. We demonstrate the ability to purify the buffer and enzyme solutions. Inorganic buffers were purified down to at least 20 fM of remaining fluorescent impurities. The exonuclease buffer solution could be cleaned down to 0.8 pM whereby its full activity was kept. Finally, we show a selection procedure for beads and present the data of a model experiment, in which immobilized DNA is degraded by an exonuclease within a polymethylmethacrylate (PMMA) microstructure. Furthermore, the mathematical processing of the obtained raw data is described. A first complete experimental cycle is shown, combining all preparatory steps which are necessary for single molecule sequencing in microstructures.

Biochemistry↗

PubMed2001

Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury.

OBJECTIVE: To evaluate the effects of PFC aerosol compared to PGI2 aerosol and NaCl aerosol on gas exchange and lung mechanics in oleic acid-induced acute lung injury. DESIGN: A prospective, controlled, randomised, in vivo animal laboratory study. SETTING: Research laboratory at an university hospital. SUBJECTS: Twenty one (n = 21) adult sheep of either gender weighing 26.8+/-6.4 kg. INTERVENTIONS: The animals were randomised to three groups: PFC aerosol (perfluorooctane), PFC group; prostacyclin aerosol (Flolan), PGI2 group; and NaCl aerosol (0.9% sodium chloride solution), control group. After induction of anaesthesia and placement of vascular catheters, lung injury was induced with 0.12 ml x kg(-1) oleic acid. Aerosols were continuously administered for 2 h using a jet nebuliser. Gas exchange, pulmonary mechanic, and haemodynamic parameters were obtained at regular intervals. MEASUREMENTS AND MAIN RESULTS: PFC aerosol increased oxygenation (PaO2) 15 min after the initiation of treatment up to 120 min (P < 0.05). Transpulmonary shunt improved in the PFC group (P < 0.05) while it did not change in the two other groups. PFC aerosol reduced maximum airway pressure (Pmax) (median) significantly from (median) 38 mbar to 32 mbar (P < 0.05). Static compliance improved significantly in the PFC group (P < 0.05). CONCLUSION: The inhalation of a PFC aerosol led to a significant improvement in pulmonary mechanics and gas exchange, which was not observed in the other two groups. These data suggest that a small dose of perfluorocarbon will have beneficial effects on gas exchange and respiratory mechanics. Therefore, the non-invasive aerosol application technique seems to be a reasonable alternative to administer perfluorocarbons in severe lung injury.

Aerosols↗

PubMed2001

Idiopathic neuritis--reasons for surgical treatment.

Idiopathic neuritis is a neurological condition of unknown etiology. Typical clinical findings are sudden onset of pain lasting for two-three weeks and remaining weakness in distal muscles. Non-surgical treatment is the consensus today for idiopathic neuritis. However, when exploring peripheral nerves affected by this syndrome that gave persistent paresis, we found an external fibrosis. After neurolysis motor function recovered. We suggest that with persisting motor deficits in idiopathic neuritis the affected peripheral nerves might be surgically explored at the latest one year after onset of the syndrome.

Adult↗

PubMed2001

Pharmacokinetics and metabolism of formestane in breast cancer patients.

Formestane (Lentaron(R), 4-hydroxyandrostenedione) is a steroidal aromatase inhibitor used for treatment of advanced breast cancer. Clinically, it is administered as a depot form once fortnightly by intramuscular (i.m.) injection. To investigate the pharmacokinetics, bioavailability and metabolism of the drug, seven patients received single 250 mg i.m. doses of commercial formestane on Days 0, 21, 35, 49 and 63 of this trial. On Day 63, three of the patients received an additional single intravenous (i.v.) pulse dose of 1 mg of 14C-labelled formestane. The plasma kinetics after i.m. dosing confirmed a sustained release of formestane from the site of injection. Within 24-48 h of the first dose, the circulating drug reached a C(max) of 48.0+/-20.9 nmol/l (mean+/-S.D.; N=7). At the end of the dosing interval, after 14 days, the plasma concentration was still at 2.3+/-1.8 nmol/l. The kinetic variables did not significantly change during prolonged treatment. Intramuscular doses appear to be fully bioavailable. Following i.v. injection of 14C-formestane, the unchanged drug disappeared rapidly from plasma, the terminal elimination half-life being 18+/-2 min (N=3). Plasma clearance, CL was 4.2+/-1.3 l/(h kg) and the terminal distribution volume V(z) was 1.8+/-0.5 l/kg. The drug is mainly eliminated by metabolism, renal excretion of metabolites accounting for 95% of dose. The excretory balance of 14C-compounds in urine and faeces totals up to 98.9+/-0.8% of the i.v. dose after 168 h. The 14C-compounds in plasma and urine were separated by HPLC, and three major metabolites were submitted to structural analysis by MS, NMR and UV spectroscopy. One of the metabolites is the direct 4-O-glucuronide of formestane. The other two represent 3-O-sulfates of the exocons 3beta,4beta-dihydroxy-5alpha-androstane-17-one and 3alpha,4beta-dihydroxy-5alpha-androstane-17-one, their ratio being 7:3. These exocons are formed by stereoselective 3-keto reduction, accompanied by reduction of the 4,5-enol function. The exocons do not inhibit human placental aromatase activity in vitro.

Aged↗

PubMed2001

Isolation of human anti-idiotypes broadly cross reactive with anti-dsDNA antibodies from patients with Systemic lupus erythematosus.

Antibodies to double stranded (ds)DNA play a central role in clinical diagnosis and disease expression in Systemic lupus erythematosus (SLE). This paper describes the isolation of anti-idiotype reagents (anti/antidsDNA) from four SLE sera and the demonstration of broad and quantitatively similar cross reactivity to both polyclonal and monoclonal anti-dsDNA antibodies isolated from SLE patients. Seven affinity-purified polyclonal and three monoclonal human anti-dsDNA preparations reacted preferentially with anti-idiotype F(ab')(2) coated plates compared to normal immunoglobulin (Ig)G F(ab')(2) coated plates in ELISA. In contrast, autoantibodies of other specificities (anti-Ro/SSA, anti-La/SSB, and anti-U(1)RNP) reacted equally with anti/anti-dsDNA F(ab')(2) and normal IgG F(ab')(2) coated plates. Such anti-idiotypic antibodies could play a significant role in the regulation of anti-dsDNA antibody levels in SLE.

Antibodies, Anti-Idiotypic↗

PubMed2001

Decoding "drug imprints" at the millennium: a proposal to increase accuracy and reduce costs.

The State of Washington mandated the use of imprints on all prescription drugs in 1980 and for "OTC's" in 1991. The FDA implemented federal requirements in 1995. Unfortunately, the FDA permitted the continued use of symbols, logotypes, and trademarks as code components, limiting the use of automated recognition systems. Analyses of several week-long samples of phone inquiries documented imprinting calls, the staff's ability to respond with an identification, the information sources used, and apparent reasons for any failure. In the first week we received 666 decoding requests, which when projected for the year amounted to > 25,000 calls. A review of 1999 data exceeded that number. Staff was able to reach drug identification in 93.8% of inquiries. Uninterpretable symbols and absence of code listings contributed to the 36 failures. Projecting over the US suggested as many as 1.25 million calls costing poison centers $25 million/y. A touch-tone telephone or website response system could permit automated responses. Neither solution is feasible without the elimination of symbols or logotypes when using an exclusively alpha-numeric code.

Drug Labeling↗

PubMed2000

The BCR/ABL tyrosine kinase induces production of reactive oxygen species in hematopoietic cells.

The BCR/ABL oncogene causes chronic myelogenous leukemia, a myeloproliferative disorder characterized by clonal expansion of hematopoietic progenitor cells and myeloid cells. It is shown here that transformation of the hematopoietic cell lines Ba/F3, 32Dcl3, and MO7e with BCR/ABL results in an increase in reactive oxygen species (ROS) compared with quiescent, untransformed cells. The increase in ROS was directly due to BCR/ABL because it was blocked by the ABL-specific tyrosine kinase inhibitor STI571. Oxidative stress through ROS is believed to have many biochemical effects, including the potential ability to inhibit protein-tyrosine phosphatases (PTPases). To understand the significance of increased production of ROS, a model system was established in which hydrogen peroxide (H(2)O(2)) was added to untransformed cells to mimic the increase in ROS induced constitutively by BCR/ABL. H(2)O(2) substantially reduced total cellular PTPase activity to a degree approximately equivalent to that of pervanadate, a well known PTPase inhibitor. Further, stimulation of untransformed cells with H(2)O(2) or pervanadate increased tyrosine phosphorylation of each of the most prominent known substrates of BCR/ABL, including c-ABL, c-CBL, SHC, and SHP-2. Treatment of the BCR/ABL-expressing cell line MO7/p210 with the reducing agents pyrrolidine dithiocarbamate or N-acetylcysteine reduced the accumulation of ROS and also decreased tyrosine phosphorylation of cellular proteins. Further, treatment of MO7e cells with H(2)O(2) or pervanadate increased the tyrosine kinase activity of c-ABL. Drugs that alter ROS metabolism or reactivate PTPases may antagonize BCR/ABL transformation.

Animals↗

PubMed2000

Functional comparison of thymic B cells and dendritic cells in vivo.

In this report we present a transgenic mouse model in which we targeted gene expression specifically to B-lymphocytes. Using the human CD19 promoter, we expressed major histocompatibility complex class II I-E molecules specifically on B cells of all tissues, but not on other cell types. If only B cells expressed I-E in a class II-deficient background, positive selection of CD4(+) T cells could not be observed. A comparison of the frequencies of I-E reactive Vbeta5(+) and Vbeta11(+) T cells shows that I-E expression on thymic B cells is sufficient to negatively select I-E reactive CD4(+) T cells partially, but not CD8(+) T cells. Thus partial negative but no positive selection events can be induced by B-lymphocytes in vivo. (Blood. 2000;95:2610-2616)

Animals↗

PubMed2000

Precursor B cell receptor-dependent B cell proliferation and differentiation does not require the bone marrow or fetal liver environment.

The capacity of precursor B (pre-B) I cells from fetal liver and bone marrow to proliferate and differentiate into surface immunoglobulin-positive immature B cells in vitro was analyzed. Both fetal liver- and bone marrow-derived progenitors do so in a pre-B cell receptor (pre-BCR)-dependent manner in tissue culture medium alone, without addition of other cells or cytokines. Approximately 20% of the initial pre-B I cells enter more than one division. Analyses at the single-cell level show that approximately 15% divide two to five times. Coculture of pre-B I cells with stromal cells did not enhance proliferation or differentiation, whereas the presence of interleukin 7, especially in combination with stromal cells, resulted mainly in the expansion of pre-B I cells and prevented their further differentiation. Thus, the environment of fetal liver or bone marrow is not required for the pre-BCR to exert its function, which is to select and expand cells that have undergone an inframe V(H)-D(H)J(H) rearrangement that produces a pre-BCR-compatible muH chain. It appears unlikely that a ligand for the pre-BCR drives this pre-B cell proliferation.

Animals↗

PubMed2000

p-Chlorophenylalanine, an inhibitor of serotonin synthesis reduces blood-brain barrier permeability, cerebral blood flow, edema formation and cell injury following trauma to the rat brain.

The role of serotonin in trauma induced alterations in blood-brain barrier (BBB) permeability, cerebral blood flow (CBF), brain edema and cell changes were examined in a new model of cortical injury in the rat using a pharmacological approach. A longitudinal incision into the right parietal cerebral cortex (about 3 mm deep and 5 mm long) was associated with a profound increase in the BBB permeability to Evans blue and [131]I-sodium, brain water content, and a reduction in the CBF in both the ipsilateral and contralateral hemispheres 5 h after trauma. Nissl staining showed a profound nerve cell reaction in the parietal cerebral cortex of both hemispheres. The intensity of these pathological changes was most pronounced in the traumatised hemisphere. Pretreatment with p-CPA, a serotonin synthesis inhibitor, significantly attenuated breakdown of the BBB permeability, brain edema and the CBF disturbances. Damaged and distorted nerve cells were markedly less frequent in p-CPA treated rats. This effect of the drug was most pronounced in the contralateral hemisphere. The observations strongly suggest that serotonin is one of the important neurochemical mediators of BBB permeability disturbances and brain edema formation in the trauma induced brain damage.

Animals↗

PubMed2000

Neurotrophic factors attenuate alterations in spinal cord evoked potentials and edema formation following trauma to the rat spinal cord.

Influence of brain derived neurotrophic factor (BDNF) and insulin like growth factor-1 (IGF-1) on spinal cord injury induced disturbances in spinal cord conduction, edema formation and cellular stress response was examined in a rat model. Pretreatment with BDNF or IGF-1 significantly attenuated the loss of SCEP negative amplitude seen immediately after spinal cord injury. In these neurotrophins treated rats, upregulation of heat shock protein (HSP 72 kD) immunoreactivity, a measure of cellular stress response and spinal cord edema formation were considerably reduced 5 h after injury. These results suggest that neurotrophic factors improve spinal cord conduction after trauma and this beneficial effect of growth factors may be related with their ability to attenuate trauma induced cellular stress response, not reported earlier.

Animals↗

PubMed2000

Growth hormone attenuates alterations in spinal cord evoked potentials and cell injury following trauma to the rat spinal cord. An experimental study using topical application of rat growth hormone.

The influence of exogenous rat growth hormone on spinal cord injury induced alterations in spinal cord evoked potentials (SCEP) and edema formation was examined in a rat model. Repeated topical application of rat growth hormone (20microl of 1microg/ml solution) applied 30min before injury and at 0min (at the time of injury), 10min, 30min, 60min, 120min, 180min, and 240min, resulted in a marked preservation of SCEP amplitude after injury. In addition, the treated traumatised cord showed significantly less edema and cell changes. These observations suggest that growth hormone has the capacity to improve spinal cord conduction and attenuate edema formation and cell injury in the cord indicating a potential therapeutic implication of this peptide in spinal cord injuries.

Animals↗

PubMed2000

Theta oscillations and the ERP old/new effect: independent phenomena?

OBJECTIVES: The hypothesis is examined whether a memory-related change in induced band power (oscillatory old/new effect) is functionally related to a memory-related increase in ERP positivity (ERP old/new effect). METHODS: In order to avoid a confounding on the measurement level, induced band power (IBP) was used as a measure that is devoid of the influence of evoked components. The EEG was recorded during a recognition memory task. RESULTS: The results show that compared to correctly rejected words, targets (remembered words) elicit a significantly larger P300. An oscillatory old/new effect was found for the delta and theta but not for the alpha band. It is manifested by an increase in delta and theta IBP which is significantly larger for targets than for correctly rejected words. It can be observed during the same time interval and shows the same topographic distribution as the ERP old/new effect. Most importantly, however, the ERP old/new effect (as well as the P300 itself) is generated by very slow frequencies which lie below the delta band. CONCLUSIONS: These findings demonstrate that the two types of old/new effects are functionally related. Possible physiological mechanisms underlying this relationship are discussed in terms of a threshold change in the cortex (generating the P300) that occurs during an increase in hippocampal theta activity (generating an increase in induced theta power).

Adult↗