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R Werner

Publications and source records attributed to R Werner.

At least 73 records · Page 4Linked to original sources

Formation of hybrid cell-cell channels.

The oocyte cell-cell channel assay was used to demonstrate that connexin-43 is a cell-cell channel-forming protein as previously shown for connexin-32. Expression of connexin-32 in one and connexin-43 in the other oocyte of a pair results in the formation of junctional conductances at rates similar to those observed when only one or the other connexin is expressed in both oocytes of a pair. This suggests that hybrid cell-cell channels form in the oocyte system. Hybrid channels also form when a connexin-43 mRNA-injected oocyte is paired with a noninjected oocyte expressing endogenous connexin. The latter hybrids have properties apparently contributed by both types of hemichannels. Pure connexin-43 channels are not voltage gated, whereas pure oocyte channels are voltage dependent; hybrids of these channels rectify.

Animals↗

Serum creatine kinase in the post-polio population.

Elevated creatine kinase (CK) levels may be noted in neuromuscular patients as a result of muscle damage and necrosis. We measured the serum CK levels of 62 post-polio patients with chronic neurologic disease (Neuro) and 13 post-polio patients with no evidence of neurologic compromise (Control). Patient groups were comparable for age, gender and years since onset of poliomyelitis. The Neuro group had substantially increased CK levels compared with the Control group, 211 +/- 19.0 IU v 114 +/- 26.2 IU (mean +/- 1 SE; P less than 0.05). Of the Neuro group, 40% had abnormally elevated CK values, compared with 8% of the Control group. Within the Neuro group, there was a robust correlation between elevated CK values and self-reports of strenuous work and level of community ambulation (P less than 0.05). These findings support the hypothesis that chronic muscle overuse may be a contributing factor to the advanced senescence reported by post-polio patients with neuromuscular compromise.

Creatine Kinase↗

Chronic tolerance to ethanol in the N:NIH rat.

As the basis for selectively breeding rats from the N:NIH stock on the basis of differential development of tolerance to ethanol, chronic tolerance to the motor-disrupting effects of ethanol was examined. Male (n = 50) and female (n = 46) rats were administered 2.5 g/kg ethanol (i.p.) and the blood-ethanol concentration (BEC) at the time of regain of aerial righting reflex (RARR) was measured. The rats were then placed in inhalation chambers for 8 consecutive days and chronically exposed to ethanol. Thirty-two hours following the cessation of chronic ethanol exposure, the rats were again given a 2.5 g/kg dose of ethanol and the BEC at RARR was again measured. The amount of chronic ethanol tolerance developed, as measured by the difference in BEC at RARR prior to and after chronic ethanol administration, was widely and normally distributed for both male and female rats. Control rats, which did not receive chronic ethanol exposure between the two tests of RARR, did not show tolerance on this measure. A negative correlation (r = -0.46) was found between BEC at RARR prior to chronic ethanol treatment and the amount of chronic tolerance developed using the difference in BEC measure. This correlation suggested that the rats who were initially more sensitive to the effects of ethanol were more likely to display the greatest amount of chronic tolerance. However, using the residuals of the regression analysis of post-chronic ethanol BEC at RARR on prechronic BEC at RARR as an index of tolerance, this negative correlation was not found.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcoholism↗

Predictive value of the three-phase technetium bone scan in diagnosis of reflex sympathetic dystrophy syndrome.

Reflex sympathetic dystrophy syndrome (RSDS) is a painful and disabling problem, the diagnosis of which can be difficult to confirm by objective measures. The three-phase technetium bone scan (TPBS), with a combined sensitivity and specificity of greater than 90%, has been recommended for use in the diagnosis of RSDS. The purpose of this study was to determine the predictive value and usefulness of the TPBS in the diagnosis of RSDS and to discover how the predictive value might be influenced by demographic and medical factors (eg, duration of symptoms). A retrospective chart review was conducted of 119 patients who underwent a TPBS as part of a workup for unexplained limb pain. Twenty-five patients met the Kozin criteria for definite or probable RSDS. All patients were injected with technetium-99m methylene diphosphonate and scanned using established criteria. The three-hour delayed image demonstrated a sensitivity of 44%, a specificity of 92%, a positive predictive value of 61%, and a negative predictive value of 86%. The blood-flow and pool-imaging phases added no further sensitivity or specificity to that achieved by the uptake scan in patients with upper-extremity involvement. Blood-flow and pool-imaging did improve the predictive value of the TPBS in patients with involvement of the lower extremities. We conclude that a more cost-effective approach to diagnosis of upper-extremity RSDS is to use the uptake scan alone.

Adult↗

Sequence of a full-length cDNA for rat lung beta-galactoside-binding protein: primary and secondary structure of the lectin.

A full-length cDNA for rat lung beta-galactoside lectin (subunit Mr approximately 14,000, lectin 14K) was cloned and the nucleotide sequence determined. The deduced amino acid sequence agrees with the amino acid composition and direct amino acid sequence analysis of purified rat lung lectin peptides. We found that the amino-terminal alanine is blocked with an acetyl group. Comparison of the amino acid sequence with other proteins shows a high degree of homology only with other vertebrate lectin sequences, supporting the suggestion that these lectins may constitute a unique class of vertebrate proteins. The amino acid composition and sequence of lectin peptides, the sequence of lectin cDNA, and isoelectric focusing of purified lectin indicate that rat lung lectin 14K is composed predominantly of a single protein. In addition, rat uterus lectin 14K was found to be the same protein as that present in lung. We characterized the secondary and tertiary structure of rat lung lectin 14K by circular dichroism, by analytical ultracentrifugation, and by computer analysis of its primary structure. Results of these experiments suggest that lectin 14K is primarily a hydrophilic protein with an asymmetric, elongated structure consisting of approximately equal amounts of alpha helix, beta sheet, beta turn, and random coil. We found that Cys-2 and Cys-130 react most rapidly with iodoacetamide; one or both of these residues may be primarily responsible for the thiol requirement of lectin activity.

Amino Acid Sequence↗

Structure of a gap junction gene: rat connexin-32.

A genomic clone for the rat liver gap junction protein (connexin-32) was isolated and characterized by restriction enzyme mapping and sequence analysis. While the complete coding sequence is contained within one uninterrupted block, the 5'-untranslated region of the transcript contains a 6.1 kb intron. Both S1 nuclease protection and primer extension assays indicate multiple transcription start sites. Sequences homologous to cAMP response elements are found near the transcription start sites and within the 3'-end of the intron.

Animals↗

Effect of metopirone-ditartrate on thermogenesis in the guinea-pig.

1. Cortisol is the major corticosteroid in the guinea-pig plasma; cortisone is detected in minor concentrations. 2. An increase in the plasma cortisol level is observed during acute cold exposure. 3. After application of metopirone-ditartrate the standard metabolic rate is depressed significantly at thermoneutrality. 4. In metopirone-treated guinea-pigs the cold-induced increase in heat production is depressed by 47% concomitant with a marked drop in body temperature. 5. It is concluded that the formation of cortisol is essential for regulatory heat production induced by acute cold exposure in the guinea-pig.

Animals↗

Tumorigenicity in the nude mouse of cocultures derived from two nontumorigenic cell types, human pituitary adenomas and mouse C3H 10T1/2 fibroblasts.

Human pituitary adenoma tissues were not tumorigenic in the hormonally manipulated nude mouse. Mouse fibroblast cells (C3H 10T1/2) also did not form tumors when inoculated alone into nude mice. When these two tissues were cocultured and coinoculated into nude mice however, the majority of inocula developed progressively enlarging tumors which could be established in tissue culture and passaged through the nude mouse. These tumors were sarcomatous histologically and thus did not resemble any human pituitary adenoma tissue injected. In order to detect any human cells in these tumors, tumor genomic DNA was subjected to Southern analysis using human repetitive Alu and HGH DNA sequence probes. Southern blot analysis of the nude mouse derived tumor genomic DNA revealed no sizeable human DNA in the mouse tumor cell genome indicating the absence of significant numbers of human cells in the tumors or the transfer of human DNA to the mouse cells. The tumors therefore arose from transformed C3H 10T1/2 cells after coculture with the human pituitary adenoma cells. These results implied that the tumorigenic transformation of susceptible C3H 10T1/2 cells in the cocultures occurred as a result of the secretion by the adenoma cells of transforming substances in the culture media or the induction of tumorigenicity through direct cell-cell contact between the two cell types.

Adenoma↗

Expression of functional cell-cell channels from cloned rat liver gap junction complementary DNA.

An oocyte expression system was used to test the relation between a complementary DNA (cDNA) clone encoding the liver gap junction protein and cell-cell channels. Total liver polyadenylated messenger RNA injected into oocytes induced cell-cell channels between paired oocytes. This induction was blocked by simultaneous injection of antisense RNA transcribed from the gap junction cDNA. Messenger RNA selected by hybridization to the cDNA clone and translated in oocyte pairs yielded a higher junctional conductance than unselected liver messenger RNA. Cell-cell channels between oocytes were also formed when the cloned cDNA was expressed under the control of a heat-shock promoter. A concentration-dependent induction of channels was observed in response to injection with in vitro transcribed gap junction messenger RNA. Thus, the liver gap junction cDNA encodes a protein that is essential for the formation of functional cell-cell channels.

Animals↗

Disintegration of a pancreatic duct stone with extracorporeal shock waves in a patient with chronic pancreatitis.

We report the case of a 33-year-old woman with chronic calcifying pancreatitis in whom an intraductal pancreatic stone with a diameter of 8 mm was successfully disintegrated with extracorporeal shock waves, permitting subsequent endoscopic extraction of the fragments. The patient had a mild attack of pancreatitis after the treatment. We conclude that shockwave lithotripsy of a pancreatic duct stone in patients with chronic pancreatitis is possible. It should, however, be viewed with reservation until further experience has been gained.

Adult↗

Correction of genetic diabetes insipidus by adult hypothalamic grafts.

Brattleboro rats manifest chronic diabetes insipidus as a result of the genetic deficiency of hypothalamic vasopressin. When basal hypothalamic tissue derived from adult F344 rats was implanted as cell suspensions or tissue blocks in the supraoptic regions of these animals, concentration of urine together with reduced urine output and water intake was observed in some animals. Histologic examination of the grafted brains from the responding animals revealed neuronal cells at the implant sites and vasopressin-staining fibers in the median eminence. This study demonstrates the feasibility of the grafting of adult cerebral tissues to correct a genetic hormonal deficiency.

Animals↗

[Determination of the phagocytosis of granulocytes by the measurement of chemiluminescence].

During two successive years the phagocytic capacity of the granulocytes of 800 children from areas with environmental pollution and with clean air was examined and compared. To this end, the luminol chemiluminescence during phagocytosis of opsonized zymosan was measured in whole blood. In all areas the phagocytic activity of the granulocytes of boys was higher than that of girls. In area with environmental pollution it was higher than in clean-air areas. The cells disposition towards phagocytosis was lower in clean-air areas than in areas with an environmental burden. Boys showed a more rapid increase in phagocytosis than girls. Between both years of examination distinct differences were observed in the level of chemiluminescence. The phagocytic defense systems of children who grew up in areas rated as having an environmental burden, showed a distinct change in activity as opposed to that of children from so-called clean-air areas. It could be proved that the phagocytic defense system of clinically healthy children reacts to environmental irritants with a change of its activity state.

Air Pollution↗

[Value of nuclear magnetic resonance tomography in the staging of tumors of the urinary bladder].

The accuracy of magnetic resonance imaging (MRI) and computed tomography (CT) in the staging of bladder tumors was evaluated in 46 patients. In the T-staging MRI was superior to CT. With MRI 38 out of the 46 tumors were correctly staged into the four groups T0, T1-T3a, T3b and T4. With CT this classification was only possible in 28 cases. The main advantages of MRI were the capability for multiplanar imaging, which improved evaluation of the bladder base and the bladder dome, as well as a better differentiation between tumor recurrency and fibrosis. In the evaluation of adenopathy MRI and CT proved to be equal and demonstrated a sensitivity of 50%.

Carcinoma, Transitional Cell↗

Translation and functional expression of cell-cell channel mRNA in Xenopus oocytes.

mRNA from estrogen-stimulated rat myometrium, a tissue known to upregulate cell-cell channels in response to this hormone, was microinjected into Xenopus laevis oocytes. The oocytes had been freed from covering layers of follicle cells and vitelline to allow direct cell membrane interactions when paired. About 4 hours after the mRNA injection, paired oocytes become electrically coupled. This coupling was due to the presence of typical cell-cell channels characterized by size-limited intercellular tracer flux, the presence of gap junctions at the oocyte-oocyte interface, and the reversible uncoupling that occurred in the presence of carbon dioxide. The induction of new cell-cell channels in the oocyte membrane was observed against a zero background or a low level of endogenous coupling, depending on the maturation stage of the oocytes. The time course of development of cell-cell coupling after the microinjection of mRNA was determined. The mRNA capable of inducing cell-cell coupling was confined to an intermediate size class when fractionated on a sucrose gradient.

Animals↗