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Biomedical subjects

A Schwartz

Publications and source records attributed to A Schwartz.

At least 235 records · Page 13Linked to original sources
PubMed1994

Functional MR mapping of activated cortical areas.

Magnetic resonance imaging (MRI) has recently been demonstrated to be sensitive to changes in neuronal activity of cortical areas. We report our initial experiences with functional MR brain mapping at high spatial resolution using a conventional whole-body MR system. A total of 10 visual and motor cortex activation studies were carried out on 8 healthy volunteers. In each examination, a time course series of 15 strongly T2*-weighted FLASH images was measured from three adjacent slices. The image analysis revealed a subtle but highly significant signal increase in cortical layers of gray matter in primary and associative visual as well as sensorimotoric cortex regions during periods of excessive brain activity provoked by photic stimuli or motoric tasks, respectively. To correlate brain structure and brain function, the computed MR brain activation maps were directly superimposed on T1-weighted anatomic spin-echo images. With this advance into the area of functional neuroimaging, MRI is moving into an established domain of positron emission tomography (PET). We, therefore, discuss the advantages and limitations of the MR method in comparison to PET as far as this can be done at present.

Adult↗
PubMed1994

Systemic distribution of beta 2-microglobulin-derived amyloidosis in patients who undergo long-term hemodialysis. Report of seven cases and review of the literature.

A search for visceral amyloid deposits was performed on autopsy material from 20 patients who had been receiving long-term hemodialysis treatment for 4 to 21 years. Visceral amyloid was found in seven patients who had undergone hemodialysis for more than 10 years. Histochemically, the amyloid was permanganate sensitive, and immunohistochemically, it stained positively for beta 2-microglobulin. The amyloid was found mainly in the wall of blood vessels, in the form of subendothelial nodules, bulging into the vessel's lumen. The amount of amyloid increased with increasing years of hemodialysis treatment. The organs most frequently involved were the heart, gastrointestinal tract, and lungs. Smaller deposits were seen in medium blood vessels of all other visceral organs. Only the spleen was "resistant" to amyloid deposition; the reason for this splenic resistance is unknown. A similar organ distribution was found in the 19 previously reported autopsy cases. Clinically, one patient experienced a perforation of the small intestine, probably related to the narrowing of the intestinal blood vessels by amyloid deposits, and this patient died of peritonitis.

Adult↗
PubMed1993

Differential contribution by conserved glutamate residues to an ion-selectivity site in the L-type Ca2+ channel pore.

In voltage-gated cation channels, it is thought that residues responsible for ion-selectivity are located within the pore-lining SS1-SS2 segments. In this study, we compared the ion permeation properties of mutant calcium channels in which highly conserved glutamate residues, located at analogous positions in the SS2 regions of all four motifs, were individually replaced. All of the mutants exhibited a loss of selectivity for divalent over monovalent cations. However, the permeation properties of the individual mutants varied in a position dependent manner. The results provide strong evidence that these glutamate residues, positioned at equivalent locations in the aligned sequences, play significantly different roles in forming the selectivity barrier of the calcium channel, and are probably arranged in an asymmetrical manner inside the ion-conducting pore.

Amino Acid Sequence↗
PubMed1993

Azidobutyryl clentiazem, a new photoactivatable diltiazem analog, labels benzothiazepine binding sites in the alpha 1 subunit of the skeletal muscle calcium channel.

[3H]Azidobutyryl clentiazem, a new photoactivable diltiazem derivative, has a higher binding affinity than azidobutyryl diltiazem. It can be covalently incorporated into the alpha 1 subunit of the skeletal muscle calcium channel by UV irradiation, which allows the benzothiazepine binding site to be determined. The photolabeled alpha 1 subunit and its proteolytic fragments were analyzed with a panel of sequence-directed antibodies. The results suggest that the linker region between segment S5 and S6 of domain IV is involved in benzothiazepine binding. This site is different from the dihydropyridine and verapamil binding sites.

Affinity Labels↗
PubMed1993

Single-channel analysis of a cloned human heart L-type Ca2+ channel alpha 1 subunit and the effects of a cardiac beta subunit.

Macroscopic and single-channel properties of a cloned human heart L-type Ca2+ channel alpha 1 subunit expressed in Xenopus oocytes were studied and the effects of a cardiac beta subunit were evaluated. The alpha 1 subunit expressed current with much slower activation and inactivation kinetics than native cells. The beta subunit increased the current amplitude and accelerated both activation and inactivation rates. Single-channel analysis revealed that the beta subunit increased the probability of channel opening and shifted the voltage-dependence to more negative potentials without affecting conductance or open time. The data suggest that the beta subunit modulates macroscopic current by increasing the probability of channel opening and shifting the voltage-dependence of the channel.

Animals↗
PubMed1993

Identification of 1,4-dihydropyridine binding domains within the primary structure of the alpha 1 subunit of the skeletal muscle L-type calcium channel.

Calcium channel blockers are drugs that bind to the alpha 1 subunit of L-type calcium channels and selectively inhibit ion movements through these channels. Determination of the mechanism of channel blockade requires localization of drug-binding sites within the primary structure of the receptor. In this study the 1,4-dihydropyridine-binding site of the membrane bound receptor has been identified. The covalently labeled receptor was purified and digested with trypsin. The labeled peptide fragments were immunoprecipitated with sequence-directed antibodies. The data indicate the existence of at least three distinct dihydropyridine-binding domains within the primary structure of the alpha 1 subunit.

Affinity Labels↗
PubMed1993

Ionic basis for OPC-8212-induced increase in action potential duration in isolated rabbit, guinea pig and human ventricular myocytes.

Changes in transmembrane ionic currents induced by OPC-8212 (3,4-dihydro-6-[4-(3,4-dimethoxybenzoyl)-1-piperazinyl]-2(1H)-quinoline) , a recently introduced positive inotropic agent which lengthens cardiac action potential duration, were examined using whole-cell voltage-clamp techniques in single rabbit, guinea pig and human ventricular myocytes. In rabbit, OPC-8212 (12 mumol/l) significantly increased membrane action potential duration measured at 90% of repolarization by an average of 88 ms (from 462 +/- 25 to 550 +/- 35 ms, n = 4; P < 0.05). In rabbit this increase in duration was not associated with significant changes in either the inward rectifier or transient outward K+ currents. The magnitude of the secondary inward current evoked from a holding potential of -50 mV was significantly increased by 97 +/- 8% (n = 6; P < 0.01) while a demonstrable delayed rectifier outward current could not be identified in the rabbit myocytes examined at room temperature. In guinea pig ventricular myocytes, where the delayed rectifier was large, 12 mumol/l OPC-8212 significantly depressed the current by 58 +/- 10% (n = 6; P < 0.01). The effects of OPC-8212 in human ventricular myocytes obtained from the explanted heart of a single patient having an idiopathic cardiomyopathy most closely resembled those observed in isolated rabbit ventricular myocytes. Thus, in rabbit and a few human ventricular myocytes examined at room temperature, OPC-8212 appeared to lengthen cardiac membrane action potential duration primarily by increasing the amplitude of the secondary inward current believed to primarily represent current through L-type Ca2+ channels. In guinea pig preparations, OPC-8212 also decreased the delayed rectifier outward K+ current which also would account for an increase in action potential duration. OPC-8212 could not be demonstrated to affect Na+ current inactivation in a manner similar to that produced by 1 mg/l veratrine, a recognized Na+ channel agonist, which dramatically slowed this process.

Action Potentials↗
PubMed1993

Molecular localization of ion selectivity sites within the pore of a human L-type cardiac calcium channel.

A highly conserved position of negatively charged amino acids is present in the SS2 segments of the S5-S6 linker regions among calcium channels. We report here that replacing Glu residues at this position alters the ion selectivity of the human cardiac calcium channel. Substituting Glu334 in motif I or Glu1086 in motif III with Lys produced mutant calcium channels that permeated sodium ions 10-fold more effectively than barium ions. More conservative changes such as substitution of Glu1086 with Gln or substitution of Glu1387 with Ala also increased sodium permeation through the mutant calcium channels. Sodium currents through the mutant calcium channels could be modulated by dihydropyridines and blocked by external divalent cations. These results suggest that Glu334, Glu1086, and Glu1387 are part of a ring of glutamate residues formed in the pore-lining SS1-SS2 region and are critical in determining ion selectively and permeability of a human cardiac calcium channel.

Amino Acid Sequence↗
PubMed1993

Detection of minor adducts in cisplatin-modified DNA by transcription footprinting.

Two DNA restriction fragments containing either a d(GC)5 or a d(TTGCTTGATTAGTTGTGTT) insert were subjected to reaction with cis-diamminedichloroplatinum(II) and were then used as templates for RNA synthesis by T7 RNA polymerase. Within the d(GC)5 insert, interstrand cross-links are preferentially formed. Within the second insert, the reactivity order of the potential binding sites is d(ApG) > d(GpC/GpC) = d(GpA) > d(GpTpG). In the presence of cyanide ions, the adducts are much less stable at the d(GpA) sites than at the d(GpCpG) sites, in double-stranded DNA.

Base Sequence↗
PubMed1993

Development of clinical standards for flow cytometry.

Flow cytometers are instruments that can determine multiparameter data simultaneously and have a great potential in providing unique information about cells. To transfer this potential to the clinical laboratory, the development and the proper use of standards and calibrators are required to ensure comparability and reproducibility of data from different flow cytometers. Although there are a number of types of standards for flow cytometry, each has its specifically designed purpose to ensure that data from this complex instrument are accurate, precise, and reproducible among instruments over time.

CD3 Complex↗
PubMed1993

Chemotactic stimulation of aggregation-stage Dictyostelium cells induces rapid changes in energy metabolism, as measured by succinic thiokinase phosphorylation.

In crude mitochondrial fractions of the cellular slime mold Dictyostelium discoideum, a 38-kDa protein can be detected in phosphorylation assays under autophosphorylation conditions in SDS polyacrylamide gels. p38 can be phosphorylated in vitro using either ATP or GTP as phosphoryl donors. After stimulation of aggregation competent cells with the chemoattractant cAMP, p38 phosphorylation pattern changes rapidly. Caffeine, a known inhibitor of cAMP relay in D. discoideum inhibits cAMP induced changes in p38 phosphorylation. The rapid changes in p38 phosphorylation after cAMP stimulation reflect changes in energy metabolism and these changes are most likely mediated by changes in internal calcium concentrations. The mitochondrial localization and other data presented on the characterization of this protein led us to the conclusion that p38 is the alpha subunit of succinic thiokinase. Data showing a correlation between in-vitro p38 phosphorylation and the metabolic state of the cells at the moment of the cell lysis are included.

Animals↗
PubMed1993

Characterization of beta subunit modulation of a rabbit cardiac L-type Ca2+ channel alpha 1 subunit as expressed in mouse L cells.

Functional properties of a rabbit cardiac alpha 1 Ca2+ channel subunit (CARD alpha 1) were investigated using the patch-clamp technique in mouse L cells, a recipient cell line which is devoid of any Ca2+ channel subunits. Cell lines resulting from stable transfection of the CARD alpha 1 subunit as well as in coexpression with a beta subunit (CARD alpha 1 beta) derived from skeletal muscle (SKM beta) were characterized. The results show that while the CARD alpha 1-Ca2+ channel activity is negligible, the Ba2+ current density is dramatically increased in the presence of beta subunit (approximately 20-fold). CARD alpha 1- and CARD alpha 1 beta-Ba2+ currents were both sensitive to the 1,4-dihydropyridine (DHP) agonist, Bay K 8644 (5- to 8-fold increase). Activation kinetics of CARD alpha 1- and CARD alpha 1 beta-Ba2+ currents were comparable. The inactivation time-course was faster (3- to 4-fold) for CARD alpha 1 beta-Ba2+ currents. We conclude that the main role of the beta subunit in heart is to modulate the L-type current density and present several lines of evidence that SKM alpha 1 and CARD alpha 1 are differentially regulated by the beta subunit.

Animals↗
PubMed1993

The gene coding for the alpha 1 subunit of the skeletal dihydropyridine receptor (Cchl1a3 = mdg) maps to mouse chromosome 1 and human 1q32.

Using both chromosomal in situ hybridization and molecular techniques, we report the genetic localization of the gene coding for the alpha 1 subunit of the skeletal slow Ca2+ current channel/DHP receptor gene (Cchl1a3) on human Chromosome (Chr) 1 (1q31-1q32 region) and on mouse Chr 1 (region (F-G)). On the basis of single-strand conformation polymorphism (SSCP-PCR) analysis in an interspecific backcross, we have determined that the Cchl1a3 = mdg (muscular dysgenesis) locus is very closely linked to the myogenin (Myog) locus.

Animals↗
PubMed1993

Molecular localization of regions in the L-type calcium channel critical for dihydropyridine action.

Sensitivity to dihydropyridines (DHPs) is a distinct characteristic that differentiates L-type Ca2+ channels from T-, N-, and P-type Ca2+ channels. To identify regions necessary for the functional effects of DHPs, chimeric Ca2+ channels were constructed in which portions of motif III or motif IV of a DHP-insensitive brain Ca2+ channel, BI-2, were introduced into the DHP-sensitive cardiac L-type Ca2+ channel. The resultant chimeric Ca2+ channels were expressed in Xenopus oocytes, and the effects of a DHP agonist and antagonist were studied. The results show that the linker region between S5 and S6 in motif IV of the L-type Ca2+ channel is a major site for DHP action. The DHP agonist and antagonist molecules interact with distinct sites on the alpha 1 subunit of the L-type Ca2+ channel. The data further show that the SS2-S6 region of motif III is not involved in DHP action but may be an important structural component of inactivation.

Amino Acid Sequence↗
PubMed1993

Inhibition of HIV-1 latency reactivation by dehydroepiandrosterone (DHEA) and an analog of DHEA.

The initial infection with human immunodeficiency virus type 1 (HIV-1) in most individuals usually results in the establishment of a latent or chronic infection before eventual progression toward acquired immunodeficiency syndrome. HIV-1 can also establish a latent or persistent infection in some T cell lines that show minimal constitutive virus expression. However, activation of the T cell lines leading to enhanced HIV-1 replication can be induced by antigens, mitogens, and cytokines (tumor necrosis factor alpha [TNF-alpha], interleukin 1, and interleukin-2). Various gene products from other viruses (HTLV-1, HSV, EBV, CMV, HBV, and HHV-6) can also enhance HIV-1 long terminal repeat (LTR)-driven reporter gene activity. On the basis of these observations, it has been proposed that reactivation of latent HIV-1 harbored in chronically infected T lymphocytes, monocytes, or macrophages plays an important role in the pathogenesis of AIDS. So far, there are no drugs or therapy available that can provide protection against HIV-1 latency reactivation. ACH-2, derived from a human T cell line (CEM), is chronically infected with HIV-1, with low levels of constitutive virus expression. ACH-2 can be converted to productive infection by stimulation of the cells with 12-O-tetradecanoylphorbol-13-acetate (TPA), mitogen or cytokines (TNF-alpha), or infection with HSV. Therefore the ACH-2 cell line is a good candidate for studying the effects of drugs on HIV-1 activation. Previously, we have reported that DHEA and synthetic analogs of DHEA can be modest inhibitors of HIV-1 IIIB replication in phytohemagglutinin-stimulated peripheral blood lymphocyte cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗