In vitro transcription by isolated nuclei of Rhynchosciara americana salivary glands. Characteristics of incorporation and inhibition by alpha-amanitin

A method for the isolation of polytene nuclei from salivary glands cells of the Diptera Rhynchosciara americana is described. The stage-specific morphological pattern of the chromosome is maintained during the isolation. The isolated nuclei show two distinct RNA polymerase activities, namely I and II, characterized on the basis of ionic requirements and alpha-amanitin sensitivity. Studies of the product under the incubation conditions show that the system allows the synthesis of high-molecular weight RNA, beside a low molecular weight peak which may comprise pre-4S and 5S RNAs. Autoradiographic studies carried out in the presence or absence of the toxin alpha-amanitin showed that micronucleoli contain products of RNA polymerase type I activity (ribosomal RNA) and that the DNA puffs are engaged in alpha-amanitin sensitive RNA synthesis and thus are sites of polymerase type II activity.     

Dopamine/neuroleptic receptors in basal hypothalamus and pituitary

In order to determine whether the basal hypothalamus or the pituitary (or both) is the likely locus of action of the tuberoinfundibular (TI) dopamine neurons, these regions were examined for dopamine and neuroleptic receptors. High affinity receptors for haloperidol and dopamine were found in the rat pituitary while none were detected in rat basal hypothalamus. The relative ability of two neuroleptics, chlorpromazine and haloperidol, to displace (3H)haloperidol from the receptor in monkey pituitary is similar to that for rat striatum. The lack of receptors capable of binding (3H)haloperidol or (3H)dopamine in the basal hypothalamus strongly suggests that the TI neurons do not produce postsynaptic effects in this region. The pituitary receptors for (3H)haloperidol and (3H)dopamine have the characteristics of a functional system. The presence of neuroleptic/dopamine receptors in the pituitary and lack of such receptors in the basal hypothalamus supports the hypothesis that dopamine may act directly as a prolactin release inhibiting factor (PIF) rather than releasing PIF from adjacent nerve terminals in the median eminence.     

Halasz syndrome revealed in the newborn infant by pulmonary arterial hypertension. Study of 4 cases

In the 4 studied patients presenting with an Halasz's syndrome, the disease was complicated since the neonatal period by pulmonary arterial hypertension and heart failure. Three out of the 4 infants rapidly died, one after pneumonectomy. The fourth is surviving and benefitted solely from medical cardiotonic treatment. The pathogenesis of early pulmonary arterial hypertension in Halasz's syndrome is complex. The possibly associated cardiac malformations, the persistance of fetal obstructive hypertension due to the increase of the flow in the left pulmonary artery, the left to right shunt induced by the abnormal venous return and predominantly by the systemic blood supply to the right lung, are responsible, at various degrees, for pulmonary hypertension. Among surgical procedures to be proposed, the simple ligation of abnormal arteries which take their origin from the aorta seems to be preferable to pneumonectomy.     

A G protein gamma subunit-like domain shared between RGS11 and other RGS proteins specifies binding to Gbeta5 subunits

Regulators of G protein signaling (RGS) proteins act as GTPase-activating proteins (GAPs) toward the alpha subunits of heterotrimeric, signal-transducing G proteins. RGS11 contains a G protein gamma subunit-like (GGL) domain between its Dishevelled/Egl-10/Pleckstrin and RGS domains. GGL domains are also found in RGS6, RGS7, RGS9, and the Caenorhabditis elegans protein EGL-10. Coexpression of RGS11 with different Gbeta subunits reveals specific interaction between RGS11 and Gbeta5. The expression of mRNA for RGS11 and Gbeta5 in human tissues overlaps. The Gbeta5/RGS11 heterodimer acts as a GAP on Galphao, apparently selectively. RGS proteins that contain GGL domains appear to act as GAPs for Galpha proteins and form complexes with specific Gbeta subunits, adding to the combinatorial complexity of G protein-mediated signaling pathways.     

Structure and stability of an immunoglobulin superfamily domain from twitchin, a muscle protein of the nematode Caenorhabditis elegans

The NMR solution structure of an immunoglobulin superfamily module of twitchin (Ig 18') has been determined and the kinetic and equilibrium folding behaviour characterised. Thirty molecular coordinates were calculated using a hybrid distance geometry-simulated annealing protocol based on 1207 distance and 48 dihedral restraints. The atomic rms distributions about the mean coordinate for the ensemble of structures is 0.55( +/- 0.09) A for backbone atoms and 1.10( +/- 0.08) A for all heavy atoms. The protein has a topology very similar to that of telokin and the titin Ig domains and thus it falls into the I set of the immunoglobulin superfamily. The close agreement between the predicted and observed structures of Ig 18' demonstrates clearly that the I set profile can be applied in the structure prediction of immunoglobulin-like domains of diverse modular proteins. Folding studies reveal that the protein has relatively low thermodynamic stability, deltaG(H2O)U-F = 4.0 kcal mol(-1) at physiological pH. Unfolding studies suggest that the protein has considerable kinetic stability, the half life of the unfolding is greater than 40 minutes in the absence of denaturant.     

The predictors of quality care

Patients' perceptions of quality are important to the performance ratings, financial viability, and operational options afforded to purchasers of health care. Through TRICARE, the Department of Defense provides care for the more than 1.5 million active duty service members charged with the defense of the nation. As patients, these service members are typically incapable of assessing the technical quality of the care they receive. This study examines the attributes of health care delivery that define military patients' perceptions of quality and discusses the implications of these assessments.     

Effect of dietary zinc levels on health and productivity of gilts and sows through two parities

The influence of 0, 50, 500 or 5,000 ppm supplemental Zn on productive characteristics, weight change, and serum and organ mineral concentrations of 60 crossbred and purebred Yorkshire gilts was evaluated. Gilts were fed their respective treatment from 30 kg body weight until the completion of two parities. Sows fed 5,000 ppm supplemental Zn weighed significantly less than sows from the other treatments when killed. Serum alkaline phosphatase activity was higher for the sows fed the highest level of Zn in all replications at 10 and 14 mo of age than for sows from the other treatments. Sows fed 0, 50 or 500 ppm had lower serum Zn and higher serum Cu concentrations than sows fed 5,000 ppm Zn at 10 and 14 mo of age. The number of pigs farrowed (total and live) and birth weight were not affected by dam's dietary treatment. However, sows receiving no additional Zn had a higher number of abnormal pigs/litter than sows on the other treatments. Sows fed 5,000 ppm additional Zn weaned fewer pigs that weighed less at weaning than sows on the other treatments. The concentration of Zn in the sow's liver increased significantly and liver Cu decreased as dietary level of Zn increased. Sows receiving 5,000 ppm Zn had lower hepatic Fe stores compared with sows receiving 500 ppm Zn. Elevated renal Cu and Zn concentrations were found in sows fed the highest level of Zn supplementation. The Zn concentration was higher and the Cu concentration lower in the aorta of sows fed 5,000 ppm Zn compared with sows fed 0 or 50 ppm additional Zn. Incidence of osteochondrosis was higher in sows supplemented with 5,000 ppm Zn than for sows from the other treatments.