Studies on the adrenal cortex of hypophysectomized rats: A model for abnormal cellular atrophy and death

Summary Biochemical and ultrastructural studies indicate that the atrophy of adrenal cortex in hypoyhysectomized rats involves the following changes: (1) One to two days after hypophysectomy, there is loss of template activity resulting from cumulative DNA-damage and heterochromatinization.In vivo ACTH-administration led to recuperation of these cells, indicating damage during hypophysectomized state to be reversible. (2) If the duration of hypophysectomy is prolonged, some of the cells become irreversibly damaged and can no longer recuperate afterin vivo ACTH administration. (3) The period of most rapid cell death is from the third to seventh day after hypophysectomy. The cause of cell death is probably due to membrane damage in the absence of protein synthesis, leading to lysis of the cells. Lysozomes and macrophages are apparently not involved.Supported by U.S.P.H.S. grants AM-5384 and AM-13724 and taken in part from dissertations submitted by Chan and by Mostafapour to Wayne State University in partial fulfillment towards the Ph.D. degree.An invited article.     

The Yale Affiliated Hospital Program in Internal Medicine

In July, 1975, the Departments of Internal Medicine at the Yale University School of Medicine and eight community hospitals in southern and western Connecticut formed the Yale Affiliated Hospital Program (YAHP) in Internal Medicine. The YAHP provides a planned and focused program of continuing education for medical staff and housestaff at the affiliated hospitals. Six formats for the over 1,000 rounds, lectures, and conferences given annually are used. The members of the YAHP also cooperate in housestaff and faculty recruiting, evaluation of quality of care and evaluation of the process of continuing medical education itself. This report summarizes the organization, goals and future plans of the YAHP.     

Evaluation of an isotope ratio method for measurement of cholesterol absorption in man

Recently an isotope ratio method (IRM) was developed for measuring cholesterol absorption in rats by analysis of radioactivity in peripheral blood (Zilversmit, D. B. 1972. Proc. Soc. Exp. Biol. Med. 140: 862-865). To validate it in man we have compared cholesterol absorption by a fecal radioactivity method (FRM) with that simultaneously measured by IRM in 14 patients (15 experiments) hospitalized on a metabolic ward. Cholesterol absorption by FRM was assayed as fecal recovery of orally administered [(14)C]cholesterol, after correction with markers for fecal flow (chromic oxide) and cholesterol degradation (beta-sitosterol). Simultaneously, [(3)H]cholesterol was administered intravenously, and the dose-normalized ratio of [(14)C]- to [(3)H]cholesterol was repeatedly assayed in plasma. After 72 hours the ratio became constant in each patient and remained so for as long as 63 weeks (five additional outpatient studies). In three patients the fecal data were unsatisfactory because of poor recoveries of chromic oxide and radioactive cholesterol. In the remaining 11 patients (12 experiments) the mean cholesterol absorption by IRM was 42.1% (range 15.7-62.9%) and by FRM 36.6% (range 13.8-58.8%). There was good to excellent agreement between the two methods in the same patient, except in one experiment. Statistical analysis of these 12 comparisons by estimating confidence intervals showed that we can be 95% confident that the two absorption methods will produce results within 5 percentage points, and 99% confident that the differences are less than 7 percentage points. Although we conclude that IRM affords results that are concordant with those obtainable by earlier validated methods, we urge that its suitability for outpatient studies be further examined in more extensive trials.     

Effects of intraventricular injection of gastrin on release of LH,prolactin, TSH and GH in conscious ovariectomized rats

Conscious ovariectomized (OVX) rats bearing a cannula implanted in the 3rd ventricle were injected with 2 μl of 0.9% NaCl containing varying doses of synthetic gastrin and plasma gonadotropin, GH and TSH levels were measured by RIA in jugular blood samples drawn through an indwelling silastic catheter. Control injections of saline iv or into the 3rd ventricle did not modify plasma hormone levels. Intraventricular injection of 1 or 5 μg gastrin produced significant suppression of plasma LH and prolactin (Prl) levels within 5 min of injection. Injection of 1 μg gastrin had no effect on plasma GH, but increasing the dose to 5 μg induced a progressive elevation, which reached peak levels at 60 min. By contrast, TSH levels were lowered by both doses of gastrin within 5 min of injection and the lowering persisted for 60 min. Intravenous injection of gastrin had no effect on plasma gonadotropin, GH and TSH, but induced an elevation in Prl levels. $\text{In}$$\text{vitro}$ incubation of hemipituitaries with gastrin failed to modify gonadotropin, GH or Prl but slightly inhibited TSH release at the highest dose of 5 μg gastrin. The results indicate that synthetic gastrin can alter pituitary hormone release in unrestrained OVX rats and implicate a hypothalamic site of action for the peptide to alter release of a gonadotropin, Prl and GH. Its effect on TSH release may be mediated both via hypothalamic neurons and by a direct action on pituitary thyrotrophs.     

Glycine cleavage and generation of one-carbon units in Euglena gracilis

Euglena gracilis Klebs (strain Z) was maintained in division synchronized autotrophic culture, receiving either air (low CO₂) or 5% CO₂ in     

The metabolic fate of triamcinolone acetonide in laboratory animals

The metabolic fate of 9-fluoro-11β,16α,17,21-tetrahydroxy-l, 4-pregnadiene-3,20-dione cyclic 16,17-acetal with 2-¹⁴C-acetone, triaacinolone acetonide (TA) was studied in rabbits, dogs, monkeys and rats and found to be qualitatively similar in all species. In the dog, rat and monkey the major excretory route was the feces irrespective of the mode of administration. In the rabbit the excreted radioactivity was equally distributed between urine and feces. The metabolites were isolated by preparative thin layer chroma tography, located by autoradiography, eluted and analyzed by MS, IR, UV and NMR. The major metabolites of triamcinolone acetonide (TA) were identified as the C-21 carboxylic acids of TA and of the 6β hydroxy-TA,(6β-OH-TA) and the previously identified (1,2) 6β-OH-TA. In addition MS and UV data indicate the presence of 9-fluoro-11β,16α, 17-trihydroxy-3,20-dioxo-1,4,6-pregnatrien-21-oic acid cyclic 16,17 acetal with 2-¹⁴C-acetone.     

Immunoglobulin isoantigens (allotypes) in the mouse

Murine antisera raised against allogeneic lymphoid cells often contain antibodies to IgM allotypes. Rarely, allotypic antibodies to IgM have been found after immunization withB. pertussis anti-B. pertussis conjugates. Using both types of antibodies, we have defined a new constant-region locus for both secreted and membrane-bound chains. This locus,Ig-6, is closely linked to the previously described H-chain constant-region loci (Ig-1 throughIg-5) and is subject to allelic exclusion. We have identified three alleles and four antigenic specificities ofIg-6.Authors listed alphabetically     

Vitamin D metabolism and expression in rats fed on low-calcium and low-phosphorus diets

1. Cholecalciferol, radioactively labelled with both (14)C and (3)H, was administered weekly for 7 weeks to rats that had been depleted of vitamin D for 4 weeks before repletion with the radioactive vitamin. This permitted measurement of the steady-state effect on vitamin D metabolism of low-calcium and low-phosphorus regimens, as compared with a normal mineral intake. These dietary manoeuvres were carried out during the last 3 weeks of repletion. Cholecalciferol, 25-hydroxycholecalciferol and 1,25-dihydroxycholecalciferol were determined in plasma, intestine, kidney and bone. Ca(2+)-binding-protein content was measured in intestine and kidneys of comparable animals. 2. In rats on the low-calcium diets, 1,25-dihydroxycholecalciferol concentration was elevated in plasma, bone, kidney and intestine, and intestinal Ca(2+)-binding protein was increased to over twice the concentration found in the control animals. 3. The low-phosphorus regimens led to a decrease in plasma phosphate and 1,25-dihydroxycholecalciferol in all tissues studied, for the latter to the point where it was undetectable in plasma and bone. Intestinal and renal concentrations of Ca(2+)-binding protein were unchanged in the low-phosphate-intake group and decreased in the very-low-phosphate-intake group. 4. It is concluded that in the rat, unlike in the chick, hypophosphataemia is not associated with a stimulation of the production of 1,25-dihydroxycholecalciferol or its expression in the synthesis of Ca(2+)-binding protein. Therefore the plasma phosphate concentration does not appear to be directly involved in the regulation of the functional metabolism of vitamin D.     

Adherent cells in tumor immunity

The present studies explored the role of adherent cells in tumor immunity. Lymph node cells from mice bearing large tumors appeared to be maximally stimulated in vivo and incapable of further stimulation by cells of the same tumor in vitro. Removal of the adherent cell population resulted in a marked decrease in the spontaneous background activity of the remaining nonadherent cells and allowed these cells to undergo stimulation when cultured in the presence of mitomycin-blocked tumor cells. The role of the adherent cell in the maintenance of a state of continuous stimulation was further elucidated by experiments in which lymph node cell populations were reconstituted from the adherent and nonadherent subpopulations. It was also shown that adherent lymphoid cells from tumor-bearing mice, but not from normal mice, were capable of stimulating tumor-immune lymphocytes in a manner similar to intact mitomycin-blocked tumor cells.