Biomedical subjects
C Walther
Publications and source records attributed to C Walther.
The ubiquitous transactivator Zfp-38 is upregulated during spermatogenesis with differential transcription.
We describe the complete nucleotide sequence of a full length cDNA clone encoding a new mouse zinc finger protein gene, Zfp-38 and localize it on chromosome 5 by the interspecific backcross analysis. The N-terminal domain of the Zfp-38 protein (64 kDa) contains 358 amino acids and the C-terminal domain of 197 residues encodes 7 zinc fingers. We also present evidence that Zfp-38 is a strong transcriptional activator. The transactivation domain was localized in the non finger region and a fusion protein containing 112 amino acid residues from this region of the Zfp-38 and the DNA binding domain of the yeast Gal 4 protein, very efficiently transactivated the expression of a reporter CAT plasmid, harboring the Gal4 target site. By in situ hybridization and northern blotting technique, the Zfp-38 transcript can be detected at a highly elevated level during spermatogenesis. Its expression accompanies the progression from pachytene spermatocytes to round spermatids. The undifferentiated spermatogonia or the haploid elongated spermatid and the spermatozoa do not show any detectable level of the transcript. Interestingly, other tissues express low levels of a slightly shorter transcript with a different 5' end as determined by RNase protection. The presence of both a transcriptional activating domain and 7 DNA binding zinc fingers, coupled with the cell type(s) specific expression pattern, suggests that Zfp-38 has the potential to regulate transcription during spermatogenesis.
Identification of proteins resembling G-protein alpha subunits in locust muscle.
In locust skeletal muscle, FMRFamide-like peptides decrease a K+ conductance. Functional data suggest the involvement of G-proteins. For identification of G-protein alpha-subunits, membranes of locust skeletal muscle were probed with ADP-ribosylating bacterial toxins, the photoreactive GTP analog, [alpha-32P]GTP azidoanilide, and with antibodies against mammalian alpha-subunits. Multiple guanine nucleotide-binding proteins of approximately 24-95 kDa were detected. Pertussis toxin catalyzed the ADP-ribosylation of two proteins comigrating with the ADP-ribosylated alpha-subunits of the mammalian G-proteins Go and Gi. Cholera toxin promoted ADP-ribosylation of a protein comigrating with mammalian cholera toxin substrates (i.e., Gs alpha-subunits). An antibody against mammalian Go alpha-subunits detected a 54-kDa protein. Thus proteins with properties of mammalian G-protein subunits are present in insect muscle.
Hyperpolarization slowly activates a potassium current in locust skeletal muscle.
An inward current activated by hyperpolarization, IK,H, was studied under voltage clamp in locust skeletal muscle. The dependence of its reversal potential on [K+]o and its insensitivity to changes in [Na+]o indicate that the underlying conductance is a K+ conductance. The instantaneous current-voltage (I-V)-relationship exhibits outward rectification. Activation and deactivation take seconds and have complex time courses. At 10 mM [K+]o activation seems to start at a voltage greater than or equal to 20 mV more positive than the resting potential (approximately -68 mV). Ba++ blocks IK,H strongly; so do Rb+ and Cs+, the latter in a voltage dependent manner. The slow inward current bears similarities to anomalous rectifiers as well as to mixed, hyperpolarization-activated K+/Na+ currents in other tissues.
Pax: a murine multigene family of paired box-containing genes.
A murine multigene family has been identified that shares a conserved sequence motif, the paired box, with developmental control and tissue-specific genes of Drosophila. To date five murine paired box-containing genes (Pax genes) have been described and one, Pax-1, has been associated with the developmental mutant phenotype undulated. Here we describe the paired boxes of three novel Pax genes, Pax-4, Pax-5, and Pax-6. Comparison of the eight murine paired domains of the mouse, the five Drosophila paired domains, and the three human paired domains shows that they fall into six distinct classes: class I comprises Pox meso, Pax-1, and HuP48; class II paired, gooseberry-proximal, gooseberry-distal, Pax-3, Pax-7, HuP1, and HuP2; class III Pax-2, Pax-5, and Pax-8; class IV Pax-4; class V Pox neuro; and class VI Pax-6. Pax-1 and the human gene HuP48 have identical paired domains, as do Pax-3 and HuP2 as well as Pax-7 and HuP1, and are likely to represent homologous genes in mouse and man. Identical intron-exon structure and extensive sequence homology of their paired boxes suggest that several Pax genes represent paralogs. The chromosomal location of all novel Pax genes and of Pax-3 and Pax-7 has been determined and reveals that they are not clustered.
Pax-6, a murine paired box gene, is expressed in the developing CNS.
A multigene family of paired-box-containing genes (Pax genes) has been identified in the mouse. In this report, we describe the expression pattern of Pax-6 during embryogenesis and the isolation of cDNA clones spanning the entire coding region. The Pax-6 protein consists of 422 amino acids as deduced from the longest open reading frame and contains, in addition to the paired domain, a paired-type homeodomain. Beginning with day 8 of gestation, Pax-6 is expressed in discrete regions of the forebrain and the hindbrain. In the neural tube, expression is mainly confined to mitotic active cells in the ventral ventricular zone along the entire anteroposterior axis starting at day 8.5 of development. Pax-6 is also expressed in the developing eye, the pituitary and the nasal epithelium.
The murine paired box gene, Pax7, is expressed specifically during the development of the nervous and muscular system.
Eight murine paired box-containing (Pax) genes have been isolated so far. The gene described here, Pax7, contains not only a paired box, but also an octapeptide and a paired-type homeobox. As shown by Northern and in situ analysis, Pax7 is expressed from day 8 to 17 p.c. during embryogenesis. At early stages Pax7 transcripts are present in a subset of cells throughout the entire brain, but later in development expression is limited to the mesencephalon. In the developing neural tube Pax7 is restricted to the dorsal ventricular zone along the entire antero-posterior axis, suggesting a role for Pax7 in the formation of certain parts of the CNS. Additionally Pax7 expression can be followed during myogenesis from the dermamyotome of the somites to the skeletal muscle tissues.
Pax8, a murine paired box gene expressed in the developing excretory system and thyroid gland.
Several mouse genes designated 'Pax genes' contain a highly conserved DNA sequence homologous to the paired box of Drosophila. Here we describe the isolation of Pax8, a novel paired box containing clone from an 8.5 day p.c. mouse embryo cDNA library. An open reading frame of 457 amino acids (aa) contains the 128 aa paired domain near the amino terminus. Another conserved region present in some other paired box genes, the octapeptide Tyr-Ser-Ile-Asn-Gly-Leu-Leu-Gly, is located 43 aa C-terminal to the paired domain. Using an interspecies backcross system, we have mapped the Pax8 gene within the proximal portion of mouse chromosome 2 in a close linkage to the surf locus. Several developmental mutations are located in this region. In situ hybridization was used to determine the pattern of Pax8 expression during mouse embryogenesis. Pax8 is expressed transiently between 11.5 and 12.5 days of gestation along the rostrocaudal axis extending from the myelencephalon throughout the length of the neural tube, predominantly in two parallel regions on either side of the basal plate. We also detected Pax8 expression in the developing thyroid gland beginning at 10.5 days of gestation, during the thyroid evagination. In the mesonephros and metanephros the expression of Pax8 was localized to the mesenchymal condensations, which are induced by the nephric duct and ureter, respectively. These condensations develop to functional units, the nephrons, of the kidney. These data are consistent with a role for Pax8 in the induction of kidney epithelium. The embryonic expression pattern of Pax8 is compared with that of Pax2, another recently described paired box gene expressed in the developing excretory system.
[A misdiagnosed nosological entity: the pyramidal muscle syndrome].
Explore the source record for details and available documents.
[Clinically controlled, integrated study on plaque inhibition with zinc-fluoride-hexetidin].
This study investigated the effect of Zinc-Fluorid-Hexetidin comparing with positive control group (Chlorhexidin) and negative control group (experimental gingivitis). The result demonstrated that a mouthrinse containing Zinc-Fluorid-Hexetidin prevented the development of plaque without complications.
[Ergonomic mode of dental treatment using equipment combination "Ergostar 90 S". Report on clinical experience].
Regarding ergonomic principles of work the new dental unit "Ergostar 90 S" was tested during a period of four weeks to estimate its efficiency. In all technical aspects it fulfills international standards. Recommendations for improvement of some ergonomic details are given.
ADP-ribosylation is involved in the integration of foreign DNA into the mammalian cell genome.
The most commonly used DNA transfection method, which employs the calcium phosphate co-precipitation of the donor DNA, involves several discrete steps (1,2). These include the uptake of the donor DNA by the recipient cells, the transport of the DNA to the nucleus, transient expression prior to integration into the host cell genome, concatenation and integration of the transfected DNA into the host cell genome and finally the stable expression of the integrated genes (2,3). Both the concatenation and the integration of the donor DNA into the host genome involve the formation and ligation of DNA strand-breaks. In the present study we demonstrate that the nuclear enzyme, adenosine diphosphoribosyl transferase (ADPRT, E.C. 2.4.2.30), which is dependent on the presence of DNA strand breaks for its activity (4,5) and necessary for the efficient ligation of DNA strand-breaks in eukaryotic cells (4,6), is required for the integration of donor DNA into the host genome. However, ADPRT activity does not influence the uptake of DNA into the cell, its episomal maintenance or replication, nor its expression either before or after integration into the host genome. These observations strongly suggest the involvement of ADPRT activity in eukaryotic DNA recombination events.
Phencyclidine depresses transmitter release at the neuromuscular synapse of the locust.
The presynaptic effects of phencyclidine (PCP) were studied in the locust extensor tibiae muscle. The neurally evoked transmitter release is diminished in the presence of 5 x 10(-6) to 5 x 10(-5) M PCP. This is indicated (1) by an increased coefficient of variation of the excitatory junction potentials; (2) by an increased rate of failures (a) with nerve stimulation in low [Ca2+]saline and (b) with focal extracellular stimulation in normal [Ca2+]saline. The rate of spontaneous transmitter release is not affected.
FMRF-NH2-like factor from neurohaemal organ modulates neuromuscular transmission in the locust.
YGGFMRFamide, FMRFamide and related peptides potentiate transmission at locust slow motor synapses. Since immunohistochemical evidence points to neurohaemal organs (NHO) as potential sources for endogenous RFamide-like peptides we have applied extracts from the metathoracic NHO, equivalent to 1/3 NHO, to the metathoracic extensor tibiae muscle. NHO-extract depolarizes the muscle fibre and increases its membrane resistance; enhances transmitter release; and increases the amplitude of contraction and the rate of relaxation. These effects are in good qualitative and quantitative agreement with those of YGGFMRFamide (5 X 10(-8) to 10(-7) M). Octopamine which partly acts like YGGFMRFamide cannot account for the NHO-effects.
Correlations between the morphological and clinical findings in a patient recovering from secondary generalised amyloidosis with renal involvement. Light- and electron microscopic investigations on serial biopsies.
We report light- and electron microscopic findings in glomerular amyloidosis (secondary amyloidosis), which occurred after recurrent empyema of the pleura. After healing of the empyema, the clinical symptoms disappeared, over a period of eight years. During the acute stage of the disease (grade II-III amyloidosis) when the nephrotic syndrome was present, amyloid deposits were seen in the mesangium and on both sides of the basement membrane of the glomerular capillaries. Furthermore, denuded basement membrane areas showing the passage of amyloid into the urinary space, and invaginations of the podocyte by straightened amyloid fibrils were found. After clinical recovery (except for a trace of proteinuria), the renal amyloidosis had electronmicroscopically transformed from an active into an inactive or resting form, while the amount of amyloid present was almost the same. In the areas of amyloid deposits, reparative changes were observed, espcially in the area of the mesangial cells and of the podocytes. The podocytes were separated from the persisting amyloid deposits by newly formed basement membrane material.
Azidophenantridinium compounds as photoaffinity labels of cholinergic proteins.
The synthesis of diazidopropidium and diazidoethidium is described. The applicability of these compounds as photoaffinity labels for cholinergic proteins has been investigated: diazidopropidium inhibits neuromuscular transmission. This inhibition is reversible if the compound is applied in the dark but becomes irreversible after irradiation with white light. Inhibition is accompanied by a disappearance of miniature endplate potentials. Electrophysiological analysis of this effect indicates that diazidopropidium acts postsynaptically by blocking the acetylcholine receptors. At the molecular level the action of diazidopropidium and diazidoethidium on acetylcholinesterase has been investigated: both compounds appear to bind to a peripheral acetylcholine binding site of this enzyme. Binding of 125I-labeled alpha-neurotoxin from Naja naja siamensis to purified membranes from Torpedo californica electric tissue rich in acetylcholine receptors is diminished after incubation and irradiation with diazidopropidium. About half of the toxin binding sites appear to be blocked by the photoaffinity label.
Aspects of turnover and biogenesis of synaptic vesicles at locust neuromuscular junctions as revealed by zinc iodide-osmium tetroxide (ZIO) reacting with intravesicular SH-groups.
Retractor unguis nerve muscle preparations from the locust were subjected to the zinc iodide-osmium tetroxide reaction (ZIO) after pre-fixation in glutaraldehyde. Applied for 18 h at 4 degrees C in the dark, ZIO reacts at pH 4.2--4.0 fairly selectively with the matrix of synaptic vesicles. Approximately 53% of the vesicles are completely and 4% partially stained. The percentage of ZIO-positive vesicles is increased to nearly 90% and reduced to 4% or less by pretreatment with SH-protecting (dithiothreitol) or SH-blocking (N-ethylmaleimide, p-chloromercuriphenyl sulfonic acid) and SH-oxidizing (azodicarboxylic acid-bis-dimethylamide) reagents, respectively. Stimulation of the motor nerve at 20 Hz for 7 min, partially fatiguing synaptic transmission, reduces the number of vesicles per square micrometer of terminal area by approximately 52%; 2 min of rest restores this number of its pre-stimulation level. These changes are chiefly accounted for by changes in the number of completely ZIO-positive vesicles. 2 min after the end of stimulation, partially ZIO-positive vesicles are three times more frequent than before. With all experimental conditions, the average volume of vesicles was as follows: ZIO-negative less than partially ZIO-positive less than completely ZIO-positive. The average volume of ZIO-positive vesicles is almost unaffected by stimulation; that of ZIO-negative vesicles is decreased by 25% immediately after stimulation, increasing with subsequent rest to the initial level after 1 h. It is suggested (a) that ZIO demonstrates intravesicular protein(s) containing SH-groups and (b) that the completely ZIO-positive vesicles represent the mature ones ready to be used for transmitter release. How the ZIO reaction differentiates between different developmental stages of vesicles which could arise from the smooth endoplasmic reticulum is discussed.
[Antigenic relationships between feline infectious peritonitis (FIP) and transmissible gastroenteritis (TGE) viruses in swine].
Explore the source record for details and available documents.