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K Jensen

Publications and source records attributed to K Jensen.

At least 91 records · Page 5Linked to original sources

Activity-dependent depression of GABAergic IPSCs in cultured hippocampal neurons.

Short-term depression of monosynaptic GABAergic inhibitory postsynaptic currents (IPSCs) evoked between pairs of cultured rat hippocampal neurons was investigated using dual whole cell patch-clamp recordings. Paired stimuli applied to the GABAergic neuron resulted in paired-pulse depression (PPD) of the second IPSC (IPSC2) at interpulse intervals from 25 to 2,000 ms. CGP 55845A, but not CGP 35348, reduced PPD marginally. Brief paired-pulse applications of exogenous GABA indicated that postsynaptic factors made only minimal contribution to PPD of IPSCs. IPSC1 and PPD was reduced on lowering [Ca2+]o and enhanced on increasing [Ca2+]o. The potassium-channel blocker 4-aminopyridine (4-AP), which increases presynaptic Ca2+ influx, enhanced IPSC1 and PPD. Chelation of residual Ca2+ in the GABAergic boutons with EGTA-AM enhanced PPD. Stimulation of the presynaptic neuron at frequencies (f) ranging from 2.5 to 80 Hz resulted in tetanic depression of IPSCs, which declined rapidly and reached a plateau depending on f and [Ca2+]o. CGP 55845A decreased tetanic depression in the first part of the train, but this could be overcome with continued stimulation. We show that GABAergic IPSCs are robustly depressed by paired-pulse stimulation in cultured hippocampal neurons. The depression of IPSCs is mainly independent of presynaptic GABAB receptors and could be caused by depletion of releasable vesicles. Depleted synapses recover with a slow time course, depending on factors that regulate [Ca2+]i in the GABAergic boutons.

4-Aminopyridine↗

Splice variants of the nuclear dot-associated Sp100 protein contain homologies to HMG-1 and a human nuclear phosphoprotein-box motif.

Sp100 and PML are interferon-inducible proteins associated with a new class of nuclear domains (known as nuclear dots or PML bodies) which play a role in tumorigenesis, virus infections, and autoimmunity. While PML is extensively alternatively spliced, only two splice variants are known for Sp100. Here we describe the identification and characterization of several Sp100 splice variant proteins and support their existence by elucidation of the 3'-end of the Sp100 gene. Some of the splice variants contain a domain of significant sequence similarity with two previously described highly related interferon-inducible nuclear phosphoproteins as well as to suppressin and DEAF-1, which altogether define a novel protein motif, termed HNPP-box. One class of splice variants contains an almost complete and highly conserved copy of the DNA-binding high mobility group 1 protein sequence and thus represent novel HMG-box proteins. When expressed transiently, both major classes of Sp100 splice variant proteins localize in part to nuclear dots/PML bodies and in addition to different nuclear domains. Furthermore, PML was occasionally redistributed. These data indicate that alternatively spliced Sp100 proteins are expressed, differ in part in localization from Sp100, and might bind to chromatin via the HMG domain.

Alternative Splicing↗

[Babesiosis].

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Babesiosis↗

[Neurosurgical treatment of trigeminal neuralgia. A comparative study of alcohol block, neurectomy and electrocoagulation].

The objective was to assess the present condition of patients previously treated with neurosurgical procedures for trigeminal neuralgia (TN). Between 1976 and 1991, 383 patients were treated for TN at the Department of Neurosurgery, Hvidovre Hospital. The latest surgical intervention performed was radiofrequency coagulation (64%), neurectomy (18%), alcohol block (16%), trigeminal tractotomy (1%), and microvascular decompression (1%). Questionnaires were sent to 316 patients treated neurosurgically for trigeminal neuralgia during the 16 year period. After radiofrequency coagulation, neurectomy and alcohol block, 83, 51 and 42% respectively experienced a pain free postoperative period. At present 49, 17 and 18% were without pain and 33, 21 and 36% had less pain compared with the preoperative state. Sequelae were described in 65, 57 and 49% of the patients. The four most common sequelae were hypaesthesia, paraesthesia, eye complaints, and dysaesthesia. If relevant pharmacotherapy has been tried without benefit, radiofrequency coagulation may still be considered as a treatment of trigeminal neuralgia.

Adult↗

Studies on aromatic amino acid oxidase activity in ram spermatozoa: role of pyruvate as an antioxidant.

Chemical antioxidants and catalase have been shown to be ineffective in improving the motility of ram spermatozoa in a chemically-defined diluent (RSD-1). In an attempt to identify the biochemical basis of this observation, the activity of aromatic amino acid oxidase (AAAO), the enzyme responsible for generation of hydrogen peroxide in ram and bull spermatozoa, has been investigated. Ram spermatozoa contained higher levels of AAAO activity than bull spermatozoa, although the physico-chemical properties of the enzyme were generally similar in both species. Components of the medium had a marked effect upon AAAO activity. In the presence of glutamate and 3-(N-morpholino)propanesulfonic acid (MOPS), AAAO activity was decreased. Pyruvate appeared to increase AAAO activity. This was due to the ability of this substance to destroy hydrogen peroxide. Pyruvate in RSD-1 works as an effective antioxidant and therefore eliminates the need for other antioxidants in the semen diluent.

Amino Acid Oxidoreductases↗

Genetic analysis using genomic representations.

Analysis of the genetic changes in human tumors is often problematical because of the presence of normal stroma and the limited availability of pure tumor DNA. However, large amounts of highly reproducible "representations" of tumor and normal genomes can be made by PCR from nanogram amounts of restriction endonuclease cleaved DNA that has been ligated to oligonucleotide adaptors. We show here that representations are useful for many types of genetic analyses, including measuring relative gene copy number, loss of heterozygosity, and comparative genomic hybridization. Representations may be prepared even from sorted nuclei from fixed and archived tumor biopsies.

Blotting, Southern↗

The cellular clearance theory does not explain the post-dialytic small molecule rebound.

AIM: (a) To determine the normalized cellular clearance (Kcn) of urea, creatinine and phosphate in patients undergoing maintenance hemodialysis; (b) To identify the factors, particularly circulatory, which determine Kcn; (c) To evaluate whether intra-dialytic blood sampling can predict the size of the post-dialytic solute concentration rebound. METHODS: Kinetic modelling of urea, creatinine and phosphate, using a two-pool variable volume computer simulation, was performed on two occasions on 34 patients undergoing maintenance dialysis. The cellular clearance was determined (a) from the size of the rebound 50 min after the end of dialysis; (b) from a mid-dialytic blood sample. Conventional two-dimensional M-mode echocardiography and Doppler peripheral blood pressure measurement were performed. RESULTS: The model produced accurate measurements of rebound Kc for urea in 93% of measurements, creatinine in 49% and phosphate in 13%. The corresponding figures for mid-dialysis Kcn were 76%, 39% and 0%. The rebound Kcn was, for urea, 8.31 +/- 4.31 ml/kg/min, and for creatinine 4.07 +/- 2.98. The mid-dialysis Kcn was, for urea, 8.57 +/- 4.25 ml/kg/min, and for creatinine 5.06 +/- 3.36. High post-dialytic rebounds (and low Kcn values) were associated with erythropoietin use (p < 0.05) and occurrence of end-dialytic hypotension (p < 0.02). Patients treated with calcium antagonists had a significantly (p < 0.001) higher Kcn. There was no correlation between mid-dialysis and rebound Kcn. Circulatory indices had no influence on Kcn. CONCLUSIONS: The two-pool cellular clearance model is compatible with urea kinetics, but not creatinine or phosphate. It is therefore unlikely that it is the correct model for small molecule kinetics. The post-dialytic solute rebound may be partly an iatrogenic phenomenon and can be reduced by preventing post-dialytic hypotension and by calcium antagonist treatment, both of which improve regional blood flow. The size of the rebound cannot be predicted by intra-dialytic blood sampling.

Blood Urea Nitrogen↗

Reproducibility and validity of physiological parameters measured in cyclists riding on racing bikes placed on a stationary magnetic brake.

UNLABELLED: The purpose of this study was: 1) To evaluate the reproducibility of physiological parameters measured during cycle exercise, and 2) To validate the predictive capacity of physiological parameters related to endurance performance. Therefore, physiological variables were measured twice during cycling exercise in a group of seven cyclists. Each cycle exercise session was separated by one week and included progressive submaximal cycling followed by a 5-km time trial. Two and three weeks later, endurance performance was evaluated by calculating average work output during a 50-km time trial (W50km). To simulate cycle performance, the cyclists' private racing bikes were placed on a stationary magnetic brake. No differences were observed between paired physiological observations in the test-retest (P>0.05). The coefficient of variation (CV) was calculated from the differences between test-retest parameters. CV for the maximal oxygen uptake (VO2max), average work output during the 5-km trial (W5km), the calculated work intensity which increased the blood lactate concentration to 2 and 4 mmol/l (W2mM and W4mM) and W50km were 1.9, 2.7, 6.1, 5.9 and 6.3%, respectively, while the 95% confidence interval (CI) showing the expected range for variation in a retest was calculated to be 80 ml.min(-1), 9, 16, 18 and 18 W, respectively. Simple linear regression showed significant correlations between VO2max, W5km, W2mM, W4mM and W50km (r-values: range 0.83-0.98, P<0.01). IN CONCLUSION: 1) Physiological parameters can be reproduced during an exercise test procedure in cyclists riding on racing bikes placed on a stationary magnetic brake, and 2) The validity of VO2max, W5km, W2mM, and W4mM as predictive parameters of endurance was demonstrated.

Adult↗

Evidence for covalent modification of the nuclear dot-associated proteins PML and Sp100 by PIC1/SUMO-1.

PML and Sp100 proteins are associated with nuclear domains, known as nuclear dots (NDs). They were discovered in the context of leukemic transformation and as an autoantigen in primary biliary cirrhosis, respectively. Both proteins are expressed in the form of many COOH-terminally spliced variants, and their expression is enhanced by interferons (IFN). The recent finding that PIC1/SUMO-1, a small ubiquitin-like protein, is covalently linked to the RanGAP1 protein of the nuclear pore complex and also binds PML in yeast cells led us to determine whether PML is covalently modified by PIC1/SUMO-1 and whether the same is true for Sp100. We found an immune reaction of PML and Sp100 proteins with a PIC1/SUMO-1-specific monoclonal antibody by immunoblotting when using cell extracts prepared from stably transfected cells inducibly expressing one isoform of each protein as well as from nontransfected cells. In contrast, both proteins did not react when synthesized in vitro. Immunofluorescence staining showed that PIC1/SUMO-1 colocalized with Sp100 and PML in NDs except in mitotic cells, in which PML and Sp100 are dissociated. Cell fractionation and immunoblotting demonstrated that PIC1/SUMO-1 immunoreactive Sp100 in IFN-treated and untreated cells was exclusively nuclear, whereas nonmodified Sp100 was also found in the cytoplasm. Taken together, these data strongly suggest covalent modification of specific nuclear isoforms of Sp100 and PML by PIC1/SUMO-1. This modification may play a regulatory role in ND structure, composition, and function.

Animals↗

Cellular localization, expression, and structure of the nuclear dot protein 52.

Nuclear dots containing PML and Sp100 proteins (NDs) play a role in the development of acute promyelocytic leukemia, are modified after infection with various viruses, and are autoimmunogenic in patients with primary biliary cirrhosis (PBC). PML and Sp100 gene expression is strongly enhanced by interferons (IFN). Based on immunostaining with a monoclonal antibody (mAb C8A2), a third protein, nuclear dot protein 52 (NDP52), was recently localized in NDs. Here we analyzed the cellular localization, expression, and structure of NDP52 in more detail. Our NDP52-specific sera revealed mainly cytoplasmic staining but no ND pattern, neither in untreated nor in IFN-treated cells. Cells transfected with NDP52 expression vectors showed exclusively cytoplasmic staining. In subcellular fractionation experiments, NDP52 was found in cytoplasmic and nuclear fractions. Unlike as described for Sp100 and PML, NDP52 mRNA and protein levels were only marginally enhanced by IFN gamma and not enhanced at all by IFN beta. NDP52 homodimerization but no heterodimerization with Sp100 or PML could be demonstrated. None of the 93 PBC sera tested contained autoantibodies against NDP52. Finally, mAb C8A2 reacted not only with NDP52 but also with a conformation-dependent epitope on the Sp100 protein. These data imply that NDP52 forms homodimers but no heterodimers with Sp100 and PML, lacks autoantigenicity in PBC, localizes mainly in the cytoplasm, and is associated with the nucleus, but not with NDs. Finally, unlike Sp100 and PML, NDP52 expression is neither markedly enhanced nor localization detectably altered by type I and II IFNs.

Animals↗

[Sexual function in patients with disseminated sclerosis. A 5-year follow-up study].

Sexual dysfunction is known to occur in multiple sclerosis (MS). The purpose of the study is to describe the change in sexual function and symptoms in a five year follow-up study. Forty-nine patients (27 females, 22 males) with definite MS were interviewed and examined. The number of patients with sexual dysfunction increased significantly (p = 0.004) and involved females and males equally. Males usually had one or two symptoms, while females frequently had two or more symptoms. It is concluded that the risk of sexual dysfunction increases over time. Further studies concerning treatment possibilities are needed.

Adult↗

[A time study in a department of histopathology].

In the Department of Histo- and Cytopathology, Herlev University Hospital, Copenhagen, a time study was performed according to the principles known from work sampling. From 22.11-12.12.1993 laboratory technicians, pathologists and secretaries were interviewed and an activity form was filled in. Six thousand nine hundred and fifty interviews were performed and the time study showed that for all groups of personnel 40% of the time was used on specimens sent for microscopy, 5% on autopsies (excl. microscopy), 15% on teaching and research, 20% on scientific meetings, reading and administration and 20% on other activities e.g. lunch, holiday, absence owing to illness. There was little variation for the different groups of personnel. On the other hand the time used on specific laboratory procedures, e.g. serial cutting, special-, and immunostaining, varied considerably for laboratory technicians, pathologists and secretaries respectively. Time studies, especially "work sampling" yield important information for the planning of laboratory services and provide documentation also for the time not directly related to routine work.

Denmark↗

Bone acidic glycoprotein-75 self-associates to form macromolecular complexes in vitro and in vivo with the potential to sequester phosphate ions.

Monoclonal antibody HTP IV-#1 specifically recognizes a complexation-dependent neoepitope on bone acidic glycoprotein-75 (BAG-75) and a Mr = 50 kDa fragment. Complexes of BAG-75 exist in situ, as shown by immunofluorescent staining of the primary spongiosa of rat tibial metaphysis and osteosarcoma cell micromass cultures with monoclonal antibody HTP IV-#1. Incorporation of BAG-75 into complexes by newborn growth plate and calvarial tissues was confirmed with a second, anti-BAG-75 peptide antibody (#503). Newly synthesized BAG-75 immunoprecipitated from mineralizing explant cultures of bone was present entirely in large macromolecular complexes, while immunoprecipitates from monolayer cultures of osteoblastic cells were previously shown to contain only monomeric Mr = 75 kDa BAG-75 and a 50 kDa fragment. Purified BAG-75 self-associated in vitro to form large spherical aggregate structures composed of a meshwork of 10 nm diameter fibrils. These structures have the capacity to sequester large amounts of phosphate ions as evidenced by X-ray microanalysis and by the fact that purified BAG-75 preparations, even after extensive dialysis against water, retained phosphate ions in concentrations more than 1,000-fold higher than can be accounted for by exchange calculations or by electrostatic binding. The ultrastructural distribution of immunogold-labeled BAG-75 in the primary spongiosa underlying the rat growth plate is distinct from that for other acidic phosphoproteins, osteopontin and bone sialoprotein. We conclude that BAG-75 self-associates in vitro and in vivo into microfibrillar complexes which are specifically recognized by monoclonal antibody HTP IV-#1. This propensity to self-associate into macromolecular complexes is not shared with acidic phosphoproteins osteopontin and bone sialoprotein. We hypothesize that an extracellular electronegative network of macromolecular BAG-75 complexes could serve an organizational role in forming bone or as a barrier restricting local diffusion of phosphate ions.

Animals↗