Biomedical subjects

T Szabo

Publications and source records attributed to T Szabo.

At least 19 recordsLinked to original sources

PubMed2001

"Flexiball" toolkit: a modular approach to self-assembling capsules.

We report the synthesis and characterization of new, self-assembling molecular capsules. The modular strategy makes use of glycoluril building blocks available in multigram amounts combined with aromatic spacer elements. The lengthy syntheses encountered with earlier generations of capsules are avoided, and several capsules of nanometer dimensions are now accessible. Single bond attachments between spacers and glycoluril modules result in monomers as dimeric capsules that are less rigid than their earlier counterparts. The host-guest properties of the homo- and heterodimeric capsules were studied using a combination of NMR and ESI-mass spectrometry. They show a less pronounced selectivity for guests of different sizes, and their increased flexibility prevents self-assembly when no rigidifying elements are present on the central spacer unit. Some of the new capsules bear inwardly directed, secondary amide N-H protons. These can be further functionalized, as shown by their methylation to give tertiary analogues. The structures hold broader implications for the placement of functional groups on concave molecular surfaces.

Alkynes↗

PubMed2001

Functional analysis of capsaicin receptor (vanilloid receptor subtype 1) multimerization and agonist responsiveness using a dominant negative mutation.

The recently cloned vanilloid receptor subtype 1 (VR1) is a ligand-gated channel that is activated by capsaicin, protons, and heat. We have attempted to develop a dominant negative isoform by targeting several mutations of VR1 at highly conserved amino acids or at residues of potential functional importance and expressing the mutants in Chinese hamster ovary cells. Mutation of three highly conserved amino acid residues in the putative sixth transmembrane domain disrupts activation of the VR1 receptor by both capsaicin and resiniferatoxin. The vanilloid binding site in this mutant is intact, although the affinity for [(3)H]resiniferatoxin (RTX) is diminished by nearly 40-fold. Interestingly, this mutant retains a significant but diminished response to protons, supporting the existence of multiple gating mechanisms for different stimuli. The mutant appears to function by interfering with the gating induced by vanilloids rather than the expression level or permeability of the receptor. In addition, this mutant was found to function as a strong dominant negative mutation when coexpressed with wild-type VR1, providing functional evidence that the VR1 receptor forms a multimeric complex. Analysis of both current density and [(3)H]RTX affinity in cells cotransfected with different ratios of wild-type and mutant VR1 is consistent with tetrameric stoichiometry for the native capsaicin receptor.

Amino Acid Substitution↗

PubMed2001

Analysis of the native quaternary structure of vanilloid receptor 1.

Vanilloid receptor subtype 1 (VR1) is a ligand-gated channel that can be activated by capsaicin and other vanilloids as well as by protons and heat. In the present study, we have analyzed the oligomeric state of VR1. Co-immunoprecipitation of differently tagged VR1 molecules indicated that VR1 can form oligomers. Using two different heterologous VR1 expression systems as well as endogenous VR1 expressed in dorsal root ganglion cells, we analyzed oligomer formation using perfluoro-octanoic acid polyacrylamide gel electrophoresis. Results were confirmed both with chemical cross-linking agents as well as through endogenous cross-linking mediated by transglutaminase. Our results clearly show that VR1 forms multimers in each of the expression systems with a homotetramer as a predominant form. The oligomeric structure of VR1 may contribute to the complexity of VR1 pharmacology. Finally, differences in glycosylation between the systems were observed, indicating the need for caution in the use of the heterologous expression systems for analysis of VR1 properties.

Animals↗

PubMed2001

N-(3-Acyloxy-2-benzylpropyl)-N'-(4-hydroxy-3-methoxybenzyl) thiourea derivatives as potent vanilloid receptor agonists and analgesics.

A series of N-(3-acyloxy-2-benzylpropyl)-N'-(4-hydroxy-3-methoxybenzyl) thiourea derivatives were investigated as vanilloid receptor ligands in an effort to discover a novel class of analgesics. The proposed pharmacophore model of resiniferatoxin. which includes the C20 homovanillic moiety, the C3-carbonyl and the orthoester phenyl ring as key pharmacophoric groups, was utilized as a guide for drug design. The compounds were synthesized after several steps from diethylmalonate and evaluated in vitro in a receptor binding assay and in a capsaicin-activated channel assay. Additional evaluation of analgesic activity, anti-inflammatory activity and pungency was conducted in animal models by the writhing test, the ear edema assay, and the eye-wiping test, respectively. Among the new compounds, 23 and 28 were found to be the most potent receptor agonists of the series with Ki values of 19 nM and 11 nM, respectively. Their strong in vitro potencies were also reflected by an excellent analgesic profile in animal tests with ED50 values of 0.5 microg kg for 23 and 1.0 microg/kg for 28. Relative to capsaicin these compounds appear to be ca. 600 and 300 times more potent. Both 23 and 28 were found to be less pungent than capsaicin based on the eye-wiping test. However, the compounds did not show significant anti-inflammatory activity. A molecular modeling study comparing the energy-minimized structures of resiniferatoxin and 35 demonstrated a good correlation in the spatial disposition of the corresponding key pharmacophores. The thioureas described in this investigation, which were designed as simplified resiniferatoxin surrogates, represent a novel class of potent vanilloid receptor agonists endowed with potent analgesic activity and reduced pungency.

Analgesics↗

PubMed2001

N-(3-acyloxy-2-benzylpropyl)-N'-dihydroxytetrahydrobenzazepine and tetrahydroisoquinoline thiourea analogues as vanilloid receptor ligands.

The vanilloid receptor represents a promising target for drug development. Building on our previous strategies which have generated potent agonists for VR1, we now describe a series of novel N-(3-acyloxy-2-benzylpropyl)-N'-dihydroxytetrahydrobenzazepine and tetrahydroisoquinoline thiourea analogues, several of which are potent VR1 antagonists. We report here the rationale for the design, the synthesis, and the in vitro characterization of activity in assays for [(3)H]resiniferatoxin binding and (45)Ca influx using heterologously expressed rat VR1.

Animals↗

PubMed2000

Ligand-induced dynamic membrane changes and cell deletion conferred by vanilloid receptor 1.

The real time dynamics of vanilloid-induced cytotoxicity and the specific deletion of nociceptive neurons expressing the wild-type vanilloid receptor (VR1) were investigated. VR1 was C-terminally tagged with either the 27-kDa enhanced green fluorescent protein (eGFP) or a 12-amino acid epsilon-epitope. Upon exposure to resiniferatoxin, VR1eGFP- or VR1epsilon-expressing cells exhibited pharmacological responses similar to those of cells expressing the untagged VR1. Within seconds of vanilloid exposure, the intracellular free calcium ([Ca(2+)](i)) was elevated in cells expressing VR1. A functional pool of VR1 also was localized to the endoplasmic reticulum that, in the absence of extracellular calcium, also was capable of releasing calcium upon agonist treatment. Confocal imaging disclosed that resiniferatoxin treatment induced vesiculation of the mitochondria and the endoplasmic reticulum ( approximately 1 min), nuclear membrane disruption (5-10 min), and cell lysis (1-2 h). Nociceptive primary sensory neurons endogenously express VR1, and resiniferatoxin treatment induced a sudden increase in [Ca(2+)](i) and mitochondrial disruption which was cell-selective, as glia and non-VR1-expressing neurons were unaffected. Early hallmarks of cytotoxicity were followed by specific deletion of VR1-expressing cells. These data demonstrate that vanilloids disrupt vital organelles within the cell body and, if administered to sensory ganglia, may be employed to rapidly and selectively delete nociceptive neurons.

Animals↗

PubMed2000

Improvements in quantitative source preparation

Quantitative source preparation is indispensable for radionuclide standardisation. To improve the source quality, a device has been developed to accelerate the evaporation of solvents from a drop deposited on a substrate. Short drying times were reached by stirring the rotating drop with multiple jets of dry nitrogen at elevated temperature. Uniform deposits with a large number of small crystals were obtained. The source-quality was checked by micrographs and scans of autoradiographs and by the shape parameters of alpha-particle spectra.

Journal Article↗

PubMed1999

Epidural resiniferatoxin induced prolonged regional analgesia to pain.

Adequate treatment of cancer pain remains a significant clinical problem. To reduce side effects of treatment, intrathecal and epidural routes of administration have been used where appropriate to reduce the total dose of agent administered while achieving regional control. Resiniferatoxin (RTX), an ultrapotent capsaicin analog, gives long-term desensitization of nociception via C-fiber sensory neurons. We evaluate here the analgesic effect on rats of epidurally administered RTX, using latency of response to a thermal stimulus in unrestrained animals. Results were compared with those for systemically administered RTX. Vehicle or graded doses of RTX were injected subcutaneously (s.c.) or through an indwelling lumbar (L4) epidural catheter as a single dose. Both routes of application of RTX produced profound thermal analgesia, reaching a plateau within 4-6 h and showing no restoration of pain sensitivity over 7 days. Vehicle was without effect. For the epidural route, the effect was selective as expected for the targeted spinal cord region, whereas the subcutaneous administration of RTX had a generalized analgesic effect. At doses yielding a tripling of back paw withdrawal latency, epidural treatment was 25-fold more effective than the subcutaneous route of application. Consistent with the regional selectivity of the lumbar epidural route, the front paws showed no more effect than by systemic RTX treatment. Binding experiments with [3H]RTX provided further evidence of the segmental desensitization induced by epidural RTX. We conclude that epidural administration of RTX at the lumbar spinal level produces profound, long-lasting, segmental analgesia to C-fiber mediated pain in the rat.

Analgesia, Epidural↗

PubMed1999

Outcome of Pseudomonas aeruginosa exit-site and tunnel infections: a single center's experience.

We reviewed the course and outcome of all Pseudomonas aeruginosa (PA) exit-site and tunnel infections (ESI/TI) that occurred at our home peritoneal dialysis (PD) unit over a 3-year period (July 1995 to June 1998). We documented PA ESI/TI in 19 out of a total of 467 patients. Of the 19 patients, 12 having local redness and tenderness but no discharge were treated conservatively with increased frequency of dressing with or without hydrogen peroxide locally. Of the 12 cases receiving local care, 7 resolved without recurrence over 14.4 months follow-up, while the remaining 5 developed persistent ESI/TI with discharge and required treatment with antibiotics. Seven more patients who initially presented with purulent discharge also received systemic antibiotics. Only 1 of the 12 patients with PA ESI/TI treated with antibiotics resolved; the remaining 11 patients developed PA peritonitis over a 1-month to 7-month period after the initial PA ESI/TI. In 2 of these 11 patients, simultaneous PD catheter removal and replacement was attempted for the treatment of PA ESI/TI, but these patients also developed PA peritonitis 1-3 weeks after the procedure. Of the 11 patients with PA peritonitis associated with PA ESI/TI, 1 died, 6 were transferred to permanent hemodialysis, and just 4 continued PD after PD catheter replacement. Though not frequent, PA ESI/TI is still a serious complication of home PD at our unit, resulting in ESI/TI-related PA peritonitis and catheter loss in 58% of cases. Local treatment of mild PA ESI/TI (redness and induration) seems to be effective. On the other hand, patients with purulent discharge are likely to develop peritonitis and technique failure despite antibiotic therapy. Early catheter replacement can be considered in these cases.

Anti-Bacterial Agents↗

PubMed1997

FTIR study on nucleotide analogues. 1. Spectral characterization of dinucleoside methylphosphonates and dinucleoside 5'-methylenephosphonates in solution and in solid phase.

Some conformational feature of dithymidine nucleotides containing natural 3'-->5' phosphodiester, methylphosphonate, or 5'-methylenephosphonate internucleotidic linkages were probed in solution and in solid phase using FTIR spectroscopy. A high similarity of the IR spectra in the region of 1800-1250 cm-1 indicates that all the investigated compounds have similar glycosidic torsion angels and the preferred conformation of the deoxyribose rings. However, small but significant differences between the Rp and Sp diastereomers of methylphosphonate analogue 5 may suggest that the association or the hydration mode of these compounds may vary.

DNA Methylation↗

PubMed1996

Projection neurons of the mormyrid electrosensory lateral line lobe: morphology, immunohistochemistry, and synaptology.

This paper describes the morphological, immunohistochemical, and synaptic properties of projection neurons in the highly laminated medial and dorsolateral zones of the mormyrid electrosensory lateral line lobe (ELL). These structures are involved in active electrolocation, i.e., the detection and localization of objects in the nearby environment of the fish on the basis of changes in the reafferent electrosensory signal generated by the animal's own electric organ discharge. Electrosensory, corollary electromotor command-associated signals (corollary discharges), and a variety of other inputs are integrated within the ELL microcircuit. The organization of ELL projection neurons is analyzed at the light and electron microscopic levels based on Golgi impregnations, intracellular labeling, neuroanatomical tracer techniques, and gamma-aminobutyric acid (GABA), gamma-aminobutyric acid decarboxylase (GAD), and glutamate immunohistochemistry. Two main types of ELL projection neurons have been distinguished in mormyrids: large ganglionic (LG) and large fusiform (LF) cells. LG cells have a multipolar cell body (average diameter 13 microns) in the ganglionic layer, whereas LF cells have a fusiform cell body (on average, about 10 x 20 microns) in the granular layer. Apart from the location and shape of their soma, the morphological properties of these cell types are largely similar. They are glutamaterigic and project to the midbrain torus semicircularis, where their axon terminals make axodendritic synaptic contacts in the lateral nucleus. They have 6-12 apical dendrites in the molecular layer, with about 10,000 spines contacted by GABA-negative terminals and about 3,000 GABA-positive contacts on the smooth dendritic surface between the spines. Their somata and short, smooth basal dendrites, which arborize in the plexiform layer (LG cells) or in the granular layer (LF cells), are densely covered with GABA-positive, inhibitory terminals. Correlation with physiological data suggests that LG cells are I units, which are inhibited by stimulation of the center of their receptive fields, and LF cells are E units, excited by electric stimulation of the receptive field center. Comparison with the projection neurons of the ELL of gymnotiform fish, which constitute another group of active electrolocating teleosts, shows some striking differences, emphasizing the independent development of the ELL in both groups of teleosts.

Animals↗

PubMed1996

Mechanisms of triggered activity induction at the border zone of normal and abnormal cardiac tissue.

Ionic mechanisms that may be involved in inducing triggered activations at the border zone (BZTAs) of normal and abnormal Purkinje fiber segments were investigated. In a two-chamber bath, fibers were divided into a normal segment and segment treated with ethylenediaminetetraacetic acid to stimulate electrophysiologic alterations 24 hours after infarct. Interventions to normal segments included 1.8 mM lidocaine (n = 10), 3 x 10(-4) mM tetrodotoxin (n = 5), 10(-3) mM aconitine (n = 4), 3 mM cesium chloride (n = 7), 10(-2) mM verapamil (n = 4), and 6-8 mM (n = 7) of K+. Ethylenediaminetetraacetic acid (3.3 mM) prolonged action potentials and induced low diastolic potentials in the normal segment border zone. Tetrodotoxin, lidocaine, and high K+ levels suppressed BZTAs; cesium chloride and aconitine increased BZTAs; and verapamil did not reduce BZTAs. The finding that BZTAs were not abolished by verapamil suggests that abnormal automaticity is not a mechanism. Apparently, BZTAs depend on the Na+ inward current activated by depolarization of the membrane secondary to depolarization of adjacent cells.

Action Potentials↗

PubMed1996

Nonoperative management of Types II and III odontoid fractures: the Philadelphia collar versus the halo vest.

The nonoperative management of patients with Types II and III fractures of the odontoid process consists of a prolonged course of cervical immobilization. The need for rigid fixation, demonstrated by the routine use of the halo vest in many institutions, has never been rigorously substantiated. We retrospectively analyzed our results with the nonsurgical management of odontoid fractures to ascertain whether cranial fixation affected overall outcome. Fifty-four patients managed at the University of Virginia Health Sciences Center, Charlottesville, VA, between 1976 and 1994 were studied. All 18 patients with Type III fractures (5 treated in the collar, 18 in the halo vest) demonstrated fracture healing and late stability. Among 36 individuals with Type II fractures, 20 were treated in the halo vest and 16 were managed in the Philadelphia collar or similar orthoses. The overall rate of late surgical intervention, the stability to flexion and extension, and the rate of bony fracture healing were not statistically different between the methods of immobilization. The rate of bony union was not significantly higher in the halo vest group (74 versus 53%), even though patients managed in the Philadelphia collar were significantly older than those in the halo vest (mean, 68 versus 44 yr). In general, nonsurgical management of Type III odontoid fractures was recommended, accompanied by use of a cervical orthosis. The determination of operative versus nonoperative treatment for Type II fractures was made on the basis of fracture anatomy, patient age, other associated injuries, and patient preference. The lack of a significant difference in the need for late surgical procedures or late instability, improved patient comfort with the cervical orthosis, and elimination of the risk of halo-related complications favored the use of the rigid cervical orthosis in the majority of these cases.

Aged↗

PubMed1993

5'-Hydrogenphosphonates of anti-HIV nucleoside analogues revisited: controversial mode of action.

The monomeric and symmetrical dimeric 5'-hydrogenphosphonate derivatives of AZT were prepared and evaluated for their inhibitory properties against HIV-1 in several cell lines. The synthesis of the compounds was achieved by reaction of AZT with in situ prepared phosphorus tris(imidazolide) or with phosphonic acid in the presence of pivaloyl chloride. The two title compounds showed in vitro anti-HIV activity similar to (but not better than) that of AZT in three cell lines which were not deficient in thymidine kinase. On the other hand they were inactive in CEM-TK- cells. Pharmacokinetic studies in several media corroborate the assumption that these compounds must not be considered as 'true antiviral agents', but that they act by releasing their nucleoside entity.

Cell Line↗

PubMed1992

Central projections and motor nuclei of the facial, glossopharyngeal, and vagus nerves in the mormyrid fish Gnathonemus petersii.

Most of the information about the anatomy of the fish's cranial nerves was collected in the first two decades of this century. Experimental analysis of the VIIth, IXth, and Xth cranial nerves by modern tract tracing techniques started about 20 years ago. Several species have been investigated to date, including one species of Agnatha (Myxinoidea), two species of elasmobranchs, and species of some orders of Teleostei like Cyprinidae, Siluriformes, Perciformes, and Gadidae. The sensory and motor nuclei of the VIIth, IXth, and Xth cranial nerves of Gnathonemus petersii were studied by anterograde and retrograde axoplasmatic transport of horseradish peroxidase and cobaltous lysine complex. The sensory nuclei form a continuous column of cells in the brain stem extending caudal to the obex. The rostral one-fourth of this column is occupied by the overlapping terminals of the VIIth and IXth nerves. The vagus nerve has 5 roots. The first 4 of these innervate the gills and the fifth supplies viscera. Afferents from the gills terminate ipsilaterally rostral to the obex in topographic order and their terminal fields overlap. Viscerosensory fibers terminate ipsilaterally in the obex region and bilaterally in the commissural nucleus of Cajal. The facial motor nucleus is located rostral to the sensory nucleus. Facial motoneurons have pear-shaped and multipolar perikarya. Their axons form a rostrally directed knee before leaving the brain. The motoneurons of the IXth and Xth nerves have a common cell column. The vagal motoneurons form a periventricular, a medial, and an intermediate cell group rostral to the obex. In the obex region and also caudal to it, a lateral and a caudal group can be distinguished. Vagal motoneurons show a topographic arrangement that is similar to that of the sensory vagal projections. The majority of motoneurons have pear-shaped perikary and ventrolaterally oriented dendrites. In the caudal nucleus the dendrites extend dorsally and overlap the terminals of sensory fibers. The axons form a dorsolaterally directed arch before joining the sensory roots. Since G. petersii uses its electrosensory system primarily for detection of food, its gustatory system is less developed than in other fishes, which possess a large number of taste buds.

Animals↗

PubMed1992

A prospective randomized study of 4.1 French catheters utilizing the percutaneous right brachial approach for the diagnosis of coronary artery disease.

To determine the utility of 4.1 French (F) catheters in diagnosing coronary artery disease, 50 patients were randomized to 4.1F Multipurpose or Judkins catheters utilizing the percutaneous right brachial approach. The randomized 4.1F tip shape catheter completed the procedure in 40% of the patients, and overall the 4.1F catheters completed the catheterization in 72% of the cases. With excessive or prolonged manipulation, the catheters were noted to kink and soften and required replacement for a catheter of similar or larger size. In 28% of the cases, larger F sizes (5F and 6F) were used to complete the procedure. There was 1 (2%) procedural complication. The total procedural success (accounting for all F sizes) without clinical complication was 98%. There was no difference between the 4.1F Judkins or Multipurpose catheter shapes for coronary arteriography by either qualitative or quantitative angiographic analysis. Left ventricular opacification was improved with the 4.1F Pigtail vs. the 4.1 Multipurpose by qualitative angiographic analysis. When the 4.1F angiograms were compared by quantitative angiography in blinded, but not randomized fashion to angiograms performed with 6F Judkins catheters, there was improved opacification of the LAD and diastolic frame of the left ventriculogram with the larger catheter. This difference was not noted with qualitative angiography. This study indicates that 4.1F catheters can be utilized from the right brachial approach for the diagnosis of coronary artery disease, thus avoiding the need for supine bedrest associated with routine femoral artery catheterization while maintaining diagnostic accuracy.

Brachial Artery↗

PubMed1992

The central connections of the olfactory bulbs in cod, Gadus morhua L.

The central projections of the "classical" olfactory system of the cod, Gadus morhua were examined with horseradish peroxidase and cobalt tracing techniques. Label was applied to the olfactory bulb or selectively to central stumps of sectioned individual olfactory tract bundlets. The olfactory bulb projects bilaterally to restricted areas of the dorsolateral, ventromedial and basolateral telencephalon, anterior commissural and preoptic areas, habenular nuclei, dorsal thalamus and to the nucleus posterior tuberis in the diencephalon. An interbulbar connection courses in the medial olfactory tract (MOT). Contralateral projections were less pronounced than on the ipsilateral side. More specifically, the lateral olfactory tract (LOT) projects ipsilaterally to the telencephalon into the Dlv, Dc, Vs and Dp areas. The lateral bundlet of the medial olfactory tract (IMOT) terminates in the Dlv and Dc areas. The medial bundlet of the medial olfactory tract (mMOT) terminates in Vv and Vd. The fused lMOT and mMOT project to the caudal telencephalon in the Vs and Dp. Neurons projecting to the olfactory bulb were located bilaterally in the telencephalon. The majority of the bulbopetal fibers course via the lateral part of the MOT; a few neurons also project to the bulb through the other bundlets of the olfactory tract. The results are compared with previous studies on the olfactory projections of other teleost species and discussed with respect to the reported functional differentiation of the olfactory system in teleosts.

Animals↗

PubMed1991

Reciprocal connections between the 'nucleus rotundus' and the dorsal lateral telencephalon in the weakly electric fish Gnathonemus petersii.

Injections of horseradish peroxidase into the dorsal lateral telencephalon of the weakly electric mormyrid fish Gnathonemus petersii resulted in retrogradely filled cells and anterogradely labelled terminals in the 'nucleus rotundus' of the ipsilateral rostral diencephalon. This connection courses via the lateral part of the lateral forebrain bundle. The present results suggest a particularly close relationship between the Dla cell group of the lateral telencephalon and the 'nucleus rotundus'.

Animals↗