Antagonistic interactions between yeast chaperones Hsp104 and Hsp70 in prion curing

The maintenance of [PSI], a prion-like form of the yeast release factor Sup35, requires a specific concentration of the chaperone protein Hsp104: either deletion or overexpression of Hsp104 will cure cells of [PSI]. A major puzzle of these studies was that overexpression of Hsp104 alone, from a heterologous promoter, cures cells of [PSI] very efficiently, yet the natural induction of Hsp104 with heat shock, stationary-phase growth, or sporulation does not. These observations pointed to a mechanism for protecting the genetic information carried by the [PSI] element from vicissitudes of the environment. Here, we show that simultaneous overexpression of Ssa1, a protein of the Hsp70 family, protects [PSI] from curing by overexpression of Hsp104. Ssa1 protein belongs to the Ssa subfamily, members of which are normally induced with Hsp104 during heat shock, stationary-phase growth, and sporulation. At the molecular level, excess Ssa1 prevents a shift of Sup35 protein from the insoluble (prion) to the soluble (cellular) state in the presence of excess Hsp104. Overexpression of Ssa1 also increases nonsense suppression by [PSI] when Hsp104 is expressed at its normal level. In contrast, hsp104 deletion strains lose [PSI] even in the presence of overproduced Ssa1. Overproduction of the unrelated chaperone protein Hsp82 (Hsp90) neither cured [PSI] nor antagonized the [PSI]-curing effect of overproduced Hsp104. Our results suggest it is the interplay between Hsp104 and Hsp70 that allows the maintenance of [PSI] under natural growth conditions.     

Sigma receptor ligands and imidazoline secretagogues mediate their insulin secretory effects by activating distinct receptor systems in isolated islets

The effects of two potent sigma receptor agonists (+)-3-PPP ((R)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine) and DTG (N,N'-di-(o-tolyl)guanidine) on the insulin secretory responses in rat islets of Langerhans were investigated. Both sigma receptor ligands were able to potentiate the insulin secretory response of islets incubated at 6 mM glucose, in a dose-dependent manner and were also able to reverse the effects of diazoxide on insulin release. When islets were treated with efaroxan, a well-characterised imidazoline insulin secretagogue, and either (+)-3-PPP or DTG together, there was an unexpected and profound absence of stimulation of insulin release as compared to when islets were incubated with each compound alone. Experiments performed with islets where there was desensitization of DTG/sigma receptor or efaroxan/imidazoline binding site mediated responses suggest that at least two distinct receptor systems appear to be involved. The complex interactions of these two classes of drug require further investigation.     

Anatomic orthotopic heart transplantation: the surgical characteristics and the advantages of the method

Standard technique of establishment of interatrial anastomoses allows to carry out orthotopic transplantation of the heart quickly and reliably. However the accumulation of collective experience evidences about a number of unavoidable shortcomings of such operation: rhythm disturbances, worsening of hemodynamics in discordant atrial contractions of recipient and donor, atrio-ventricular valve insufficiency, thromboembolism, coronary fistulas. Since 1990 in RRCS 25 operations were carried out with the use of standard technique and we also met with the risk of developing the above complications. In experiment on 30 mongrel dogs the technique of anatomical, truly orthotopic transplantation of the heart with six anastomoses was developed: left pulmonary veins with a common cuff, separate anastomoses of the superior and inferior caval veins, anastomoses of aorta and pulmonary artery. Particular attention was paid to developing of original surgical modes for prophylaxis of stenoses in the area of anastomoses of pulmonary and caval veins. In 1997 the anatomical technique of heart transplantation was successfully introduced by us into clinical practice. Clinical electrophysiological, echocardiographic and functional examinations have confirmed the results of the experiments and have evidenced for substantial advantages of the anatomical technique of orthotopic transplantation of the heart. By reliability the new operation is not inferior to standard method of N. Shumway.     

Effect of dietary monensin on ovarian response following gonadotropin treatment in prepuberal heifers

Twenty prepuberal Charolais X Brahman-Hereford heifers were randomly assigned to be fed a concentrate containing either 0 mg (C) or 200 mg (M) monensin sodium/head/day. Coastal bermudagrass hay was fed ad libitum. Average daily gain was similar for the two groups. Each heifer received 1 mg of porcine follicle stimulating hormone (FSH-P) (Armour) at 0800 and 2000 hr on days 22 through 26 (10 mg total) and 2,500 IU human chorionic gonadotropin (HCG) on day 27. Flank laparotomy was performed on day 30, for examination of ovaries, and ovariectomy was performed on day 37. The average ovarian size +/- standard error at day 15 ws 3,730 +/- 66 mm3 and 1,848 +/- 55 mm3 for groups M and C, respectively (P < .025), as measured by rectal palpation. Numbers of ovulation sites measured on day 30 were 9.1 +/- 2.2 and 4.9 +/- 1.8 per heifer for groups M and C, respectively (P < .01). After ovariectomy on day 37, heifers fed M were found to have greater ovarian weight (P < .05), more corpora lutea (CL) (P < .05), greater total luteal weight (P < .05), more follicles (P < .01) and greater weight of follicular fluid (P < .05) and stroma (P < .025) than controls. CL were analyzed for progesterone content by spectrophotometric procedures. Heifers fed M had slightly larger CL (P < .10) with progesterone concentrations similar to those in CL from controls. This resulted in more luteal progesterone per CL and more luteal progesterone per heifer in the M heifers than in the controls. Prepuberal heifers fed M, which caused the expected shifts in rumen fermentation and volatile fatty acid production, exhibited an enhanced ovarian response to gonadotropins compared to that exhibited by controls.