Dantrolene prevents the malignant hyperthermic syndrome by reducing free intracellular calcium concentration in skeletal muscle of susceptible swine

Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle triggered when susceptible subjects are exposed to volatile anesthetic agents and/or depolarizing muscle relaxants. We have used Ca2+ selective microelectrodes to measure in vivo the intracellular free [Ca2+] in skeletal muscle of MH susceptible swine before and after the administration of dantrolene. We have investigated the effectiveness of this muscle relaxant in preventing clinical MH and the relationship between the resting intracellular free [Ca2+] and the probability of inducing the MH syndrome. The resting intracellular free [Ca2+] was 0.41 +/- 0.01 microM (M +/- SEM), which agrees with our previous measurements in susceptible swine. The administration of 0.5, 1, 2, 2.5 and 3 mg/Kg Dantrolene, reduced the intracellular free [Ca2+] to 0.31, 0.21, 0.09, 0.08, 0.08 microM respectively. The 0.5 mg/Kg dose induced a moderate decrease of [Ca2+]i and failed to prevent the MH syndrome after exposure to halothane (2%). The 1 mg/Kg dose produced a further reduction in [Ca2+]i and was sufficient to prevent the clinical syndrome in 2 out of 3 animals. The 2.5 mg/Kg dose was uniformly protective in all animals. These results suggest that the mechanism by which dantrolene protects susceptible animals exposed to triggering agents is by reducing the intracellular free [Ca2+] in skeletal muscle.     

Antitrypanosomal action of cis-diamminedichloroplatinum (II) analogs

The present report deals with the alterations produced by cis-diamminedichloroplatinum (II) (DDP), and 2 of its analogs: cis-Pt(II)(tranylcypromine)2Cl2 and cis-Pt(II)(benzothiazole)2Cl2 in cultured epimastigote forms of Trypanosoma cruzi. Studies have been performed at the ultrastructural level and the inhibitory effect of these complexes on macromolecule synthesis, evaluated by 3H-thymidine, 3H-uridine, and 3H-leucine incorporation, has been investigated. DDP at concentrations of 50 and 100 micrograms/ml does not inhibit significantly the incorporation of radioactive precursors, but a clear decrease was observed with the 2 analogs. Eight hours of treatment at a concentration of 10 micrograms/ml rendered in all 3 cases an increase in autophagic vacuoles and lipids as well as an abnormal condensation of the nucleus chromatin.     

Isolation and characterization of the nuclear gene encoding the Rieske iron-sulfur protein (RIP1) from Saccharomyces cerevisiae

The nuclear gene encoding the Rieske iron-sulfur protein of the cytochrome bc1 complex of the mitochondrial respiratory chain has been isolated and characterized from Saccharomyces cerevisiae. We used a segment of the iron-sulfur protein gene from Neurospora crassa (Harnisch, U., Weiss, H., and Sebald, W. (1985) Eur. J. Biochem. 149, 95-99) to detect the yeast gene by Southern analysis. Five different but overlapping clones were then isolated by probing a yeast genomic library carried on YEp 13 by colony lift hybridization. Several approaches confirmed that the isolated DNA contained the gene for the Rieske iron-sulfur protein. The yeast gene, which contains no introns, can be expressed in Escherichia coli. A 900-base pair HindIII-EcoRI fragment was subcloned into pUC19 and directed the synthesis of immunodetectable protein. The gene was also identified by disruption of its chromosomal copy by homologous integration. A 400 base pair PstI-EcoRI fragment cloned adjacent to a HIS3 marker in pUC18 was used as an integrating vector. HIS+ transformants were obtained which were unable to grow on the nonfermentable carbon source glycerol. Southern analysis of the respiration deficient (gly-) strains confirmed that the chromosomal copy of the gene was disrupted, and immunoblots of extracts of the transformants indicated a lack of iron-sulfur protein. A respiration-deficient integrant was transformed to GLY+ by a 2-kilobase pair HindIII-BglII fragment, including a complete copy of the gene, carried on a multicopy episomal vector. Immunoblots with monoclonal antibodies to the iron-sulfur protein indicated overproduction of the protein in the complemented strain and revealed expression of approximately equal amounts of mature iron-sulfur protein and of a protein approximately 3 kDa larger than the mature protein in the complemented strain. A 1.2-kilobase pair segment of DNA from the clone which complemented the disrupted strains was sequenced and found to contain an open reading frame of 645 nucleotides, capable of encoding a 21,946-dalton protein. The gene is flanked by consensus signal sequences for initiation and termination which are common in yeast and is preceded by a possible upstream activating sequence. Amino acid sequence analysis of the amino-terminal end of the mature iron-sulfur protein agreed exactly with that predicted by the nucleotide sequence starting at Lys-31.(ABSTRACT TRUNCATED AT 400 WORDS)     

Seasonal changes in the effects of arginine vasotocin and stretch on Anolis uterine contractions in vitro

We determined the effects of acute stretch on spontaneous and arginine vasotocin (AVT)-driven contractions of the Anolis carolinensis uterus in vitro. Whole uteri from reproductively inactive females (October) were placed in a bath of oxygenated 32 degrees C Anolis "Ringer's." Two initial tensions were utilized, 1.5 g or 15 g, the latter being an estimate of the tension on the wall of a uterine compartment. Uteri were then exposed to either saline or AVT (50 ng/ml), and spontaneous or AVT-driven contractions were recorded for 20 min with the use of a strain gauge and physiograph. A similar experiment was performed on uteri from reproductively active females in the summer (June). Our results indicate that the effects of acute stretch and AVT on uterine contractility were qualitatively similar in summer and fall. That is, AVT induced a tonic contraction; stretch decreased the duration of the tonic contraction; the saline-treated uteri exhibited spontaneous rhythmic contractions; AVT increased the amplitude of the rhythmic contractions, but only at the lower tension; there were no effects of AVT on the timing (contraction interval, duration, rest interval) of the rhythmic contractions; and stretch increased the frequency of the rhythmic contractions. Season greatly influenced the magnitude of these contractile phenomena. Uteri tested during the breeding season exhibited greater distensibility, an increase in the amplitude and duration of the AVT-driven tonic contraction, and an increase in the frequency of both spontaneous and AVT-driven rhythmic contractions because of a decrease in both contraction duration and rest interval.(ABSTRACT TRUNCATED AT 250 WORDS)     

Short and plump physique of Mexican-American children

Mexican-American children are shorter but relatively heavier than non-Hispanic whites and blacks. The objectives of this paper are to assess the extent to which this "short and plump" physique occurs in data collected in two national surveys (HANES I and II); to determine variations by age, sex, and socioeconomic status; and to investigate the anthropometric characteristics that may account for the overweight. Three groups, defined on the basis of reported ancestry and observed race, are studied: Mexican-Americans (MEXAME), non-Hispanic Whites, (EURAME), and blacks (BLACK). Short stature was clearly associated with the poverty index (PI) in all three groups. MEXAMEs with a PI greater than 1.6 were similar in stature to EURAMEs at the same income level at ages 1-11 years but not at 12-17 years. On the other hand, MEXAMEs were shorter than BLACKs at all ages and income levels. The body mass index (kg/cm2) and poverty were unrelated. With respect to the anthropometric characteristics examined that are related to the body mass index, MEXAMEs and EURAMEs were similar in sitting height as a proportion of total height, arm muscle and fat areas, and triceps skinfold but different in the following ways: MEXAMEs had narrower elbow but broader bitrochanteric breadths and larger chest circumferences and subscapular skinfolds. Greater upper body dimensions and fatfolds seem to best describe the physique of MEXAMEs. However, in multiple regressions, these anthropometric characteristics failed to account fully for the greater relative weight of MEXAMEs as compared to EURAMEs.     

Host preferences of two encyrtid parasitoids for the Columbian Phenacoccus spp. of cassava mealybugs

In a choice test among six life stages of Phenacoccus herreni Cox & Williams, Epidinocarsis diversicornis (Howard) used its antennae to examine adult and 3rd stadium females more than other stages and preferentially attempted to oviposit in these plus 2nd stadium females. Success of ovipositor insertion was unaffected by host stage. The outcome of these behaviors was preferential oviposition by E. diversicornis in the large female host stages. Acerophagus coccois Smith also preferentially examined larger female mealybugs (second and third stadium nymphs and adults) more than other stages and successfully inserted its ovipositor in these stages more often than in second stadium male nymphs and male cocoons, resulting in a similar preference in this species for larger female host stages. When given a choice between adult female hosts of two species, P. herreni and Phenacoccus gossypii Townsend & Cockerell, E. diversicornis exhibited a clear preference for P. herreni; whereas A. coccois preferred P. gossypii.

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Résumé Epidinocarsis diversicornis (Howard), ayant la possibilité de choisir entre six stades différents de Phenacoccus herreni Cox & Williams, examine avec ses antennes plus particulièrement les adultes et les larves femelles du 3ème stade, et essaie de pondre de préférence dans ces stades et les larves femelles de second stade. L'insertion de la tarière s'effectue aussi bien quel que soit le stade de l'hôte. Il résulte de ces différents aspects du comportement que E. diversicornis pond de préférence dans les femelles des stades les plus avancés. Acerophagus coccois Smith préfère aussi examiner les cochenilles femelles les plus grosses (second et 3ème stade larvaires et adulte), et introduit sa tarière avec succès dans ces stades plus souvent que dans les larves mâles de second stade ou les cocons mâles; il en résulte aussi pour cette espèce une préférence pour les femelles des stades les plus gros.Quand on leur a donné le choix entre des femelles des deux espèces de cochenilles (P. herreni et Phenacoccus gossypii Towsend & Cockerell), E. diversicornis manifestait une nette préférence pour P. herreni, tandis que A. coccois préférait P. gossypii.

     

Alteration of Extracellular Enzymes in Pinto Bean Leaves upon Exposure to Air Pollutants, Ozone and Sulfur Dioxide

Diamine oxidase and peroxidase, associated with the wall in pinto bean (Phaseolus vulgaris L. var Pinto) leaves, can be washed out by vacuum infiltration and assayed without grinding the leaf. The diamine oxidase activity is inhibited in vivo by exposure of the plants to ozone (dose of 0.6 microliters per liter x hour), whereas the peroxidase activity associated with the wall space is stimulated. This dose does not cause obvious necrosis or chlorosis of the leaf. These alterations are greater when the dose of ozone exposure is given as a triangular pulse (a slow rise to a peak of 0.24 microliters per liter followed by a slow fall) compared to that given as a constant square wave pulse of 0.15 microliters per liter for the same 4 hour period. Exposure of the plants to sulfur dioxide (at a concentration of 0.4 microliters per liter for 4 hours) does not result in any change in the diamine oxidase or peroxidase activities, yet the total sulfhydryl content of the leaf is increased, demonstrating the entry of sulfur dioxide. These two pollutants, with different chemical reactivities, affect the activities of the extracellular enzymes in different manners. In the case of ozone exposure, the inhibition of extracellular diamine oxidase could profoundly alter the movements of polyamines from cell to cell.     

Renin release responses to in vitro prostaglandin challenge in the summer-active ground squirrel, Spermophilus lateralis

1. Using a renal cortical slice preparation from the summer-active ground squirrel, Spermophilus lateralis, resting renin release (RR) and tissue cyclic AMP content (TcAMPC) levels were found to be significantly higher and lower, respectively, than those previously reported for its hibernating counterpart. 2. At a 10(-5) M dose, PGE2 but not PGE1, PGF2-alpha or PGA1, significantly stimulated RR in the summer-active ground squirrel (SAGS). 3. Addition of agents which normally increase TcAMPC significantly potentiated the effect of PGE1, while preventing that of PGE2, on RR and TcAMPC. 4. Opposite TcAMPC changes may mediate the in vitro RR responses to PGE1 and PGE2 administration in the SAGS.     

Regulation of reduced nitrogen assimilation in Rhodobacter capsulatus E1F1

The phototrophic bacterium Rhodobacter capsulatus E1F1 assimilates ammonia and other forms of reduced nitrogen either through the GS/GOGAT pathway or by the concerted action of l-alanine dehydrogenase and aminotransferases. These routes are light-independent and very responsive to the carbon and nitrogen sources used for cell growth. GS was most active in cells grown on nitrate or l-glutamate as nitrogen sources, whereas it was heavily adenylylated and siginificantly repressed by ammonium, glycine, l-alanine, l-aspartate, l-asparagine and l-glutamine, under which conditions specific aminotransferases were induced. GOGAT activity was kept at constitutive levels in cells grown on l-amino acids as nitrogen sources except on l-glutamine where it was significantly induced during the early phase of growth. In vitro, GOGAT activity was strongly inhibited by l-tyrosine and NADPH. In cells using l-asparagine or l-aspartate as nitrogen source, a concerted induction of l-aspartate aminotransferase and l-asparaginase was observed. Enzyme level enhancements in response to nitrogen source variation involved de novo protein synthesis and strongly correlated with the cell growth phase.Abbreviations ADH l-alanine dehydrogenase - AOAT l-alanine:2-oxoglutarate aminotransferase - Asnase l-asparaginase - GOAT Glycine: oxaloacetate aminotransferase - GOGAT Glutamate synthase - GOT l-aspartate: 2-oxoglutarate aminotransferase - GS Glutamine synthetase - HPLC High-Pressure Liquid Chromatography - MOPS 2-(N-morpholino)propanesulfonic acid - MSX l-methionine-d,l-sulfoximine     

Immunophenotyping of acute myeloid leukaemia: relevance of analysing different lineage-associated markers

The immunophenotype of 135 previously untreated patients with FAB defined acute myeloid leukaemia (AML) was studied at diagnosis. The panel of reagents included monoclonal antibodies (MoAb) recognising myeloid-associated determinants (CD11, CD13, CD14, CD33 and others) as well as MoAb directed towards lymphoid antigens (CD7, CD10, CD19) and TdT. The results indicate that CD13 and/or CD33 are consistently expressed in AML and only rarely in ALL blasts (131/135 + ve cases, versus 4/130 in ALL). Lymphoid antigen expression was rarely detected when CD10 and CD19 were investigated in AML (0.9% and 2% + ve cases, respectively), whereas significant positivities were found for TdT and CD7 (20% and 10% respectively). Concerning FAB subtypes, two new MoAb (LAM3 and LAM7) proved very useful in the specific recognition of AML with monocytic features. The phenotype CD13+ and/or CD33+, CD9+, HLA-DR- was found to be almost exclusive for M3 AML. The response to induction chemotherapy was analysed in CD7+ and in TdT+ patients. In the latter group a statistically significant lower response rate was found with respect to TdT-ve-AML patients.