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Biomedical subjects

A Gertler

Publications and source records attributed to A Gertler.

At least 127 records · Page 7Linked to original sources
PubMed1985

The fate of [125I]-labeled human leukocyte-derived alpha interferon in the rat.

In order to follow the catabolic fate of interferon in the body, [125I]-labeled human alpha interferon was infused into rats. Interferon activity and TCA-precipitable radioactivity in the blood reached a steady state after 60 min of infusion, while TCA-soluble radioactivity continued to increase during the entire infusion period. A massive accumulation of both active interferon and degradation products of interferon were found in the kidney at the end of the infusion. Most of the interferon activity was found in the mitochondrial-lysosomal fraction, while most of the interferon degradation products were found in the supernatant fraction. Ligation of the kidney's blood vessels resulted in a large increase of interferon activity in the blood. Only a small increase was found in other organs, with no increase at all in radioactive degradation products. Our results suggest the kidney to be the main site of interferon catabolism. Low molecular weight degradation products are excreted from the kidney into the blood and are at least partially secreted in the urine and also taken up by other organs.

Animals↗
PubMed1985

Digestion of elastin by porcine pancreatic elastase I and elastase II.

Elastin was fully solubilized by digestion with elastase I or elastase II. Each digest was separated into high-molecular weight and low-molecular weight fractions that were characterized by the correspondence to their amino acid content, N-terminal sequence and C-terminal amino acids. It was found that although the relative amount of amino acids in the low-molecular weight fraction obtained by digestion with elastase I was lower than in digestion with elastase II, no major difference in the type of bonds cleaved in the low- or high-molecular weight fractions of each digest could be seen. There is, however, a remarkable difference in the type of bond cleaved by the two enzymes. While elastase I cleaves mostly Ala-Ala and also Ala-Gly bonds, elastase II hydrolyzes Leu-Ala, Leu-Gly, Phe-Ala, Phe-Gly and Tyr-Ala, Tyr-Gly bonds. Theoretical calculations led us to suggest both digests are composed of cross-linked peptides that vary not only in the molecular size but also in the number of cross-links found in peptides of the same size.

Amino Acid Sequence↗
PubMed1985

Enhancement of human growth hormone-stimulated mitogenesis of Nb2 node lymphoma cells by 12-O-tetradecanoyl-phorbol-13-acetate.

The tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) enhanced human (h) GH- and ovine PRL-stimulated mitogenesis of the Nb2-11C clone of rat lymphoma cells. Maximal enhancement of 25% in the proliferation rate was achieved with 20 nM TPA. The enhancing effect was found at all levels of hGH (0.031-2.0 ng/ml), but was more pronounced at lower hormone concentrations. TPA alone had no effect on cell proliferation, and its activity was absolutely dependent on the simultaneous presence of the lactogenic hormones. We have analyzed the changes that occurred in the distribution of cells in different phases of the cell cycle during the first 22 h after exposure to hGH and measured the proliferation rate through 3 days. We have found that the mitogenic effect of hGH resulted from 1) an increase in the rate of G0/G1----S transition, 2) a decrease in the lag period required for entry into the S phase, and 3) an increase in the number of cells entering this transition. TPA enhanced all three effects. Binding of [125I]hGH was not affected by prior exposure to TPA, suggesting that the effect of TPA is at a postreceptor level. Proliferation of an autonomous clone of Nb2 cells that does not require lactogenic hormones for growth was not stimulated by TPA, although these cells bound [3H]TPA to the same extent as the PRL-dependent Nb2-11C clone.

Animals↗
PubMed1984

Binding sites of human growth hormone and ovine and bovine prolactins in the mammary gland and the liver of lactating dairy cow.

Membrane preparations and Triton X-100 solubilized fractions from the mammary gland and liver of the lactating dairy cow were capable of specific binding of [125I]hGH and [125I]oPRL. The specific binding of the latter was significantly lower and could not be increased by higher receptor levels. Displacement studies of [125I]hGh by hGH, bPRL and oPRL revealed that the two latter hormones have a 20-40-fold lower affinity for the receptor than hGH, although strong indications exist that they all bind or the same sites. This feature is unique for cows and does not exist or is much less pronounced in rodents.

Animals↗
PubMed1983

Down-regulation of prolactin receptors in the liver, mammary gland and kidney of female virgin rat, infused with ovine prolactin or human growth hormone.

Down regulation of prolactin (PRL) receptors resulting from i.v. infusion of oPRL or human growth hormone (hGH) into female virgin rats was demonstrated. A decrease of over 85% in the number of free receptors was achieved within 15 - 30 min using infusion of oPRL or hGH at 25 micrograms/h and remained at this level until the end of infusion. Ovine growth hormone or recombinant bovine growth hormone at ten-fold higher concentration had no effect at all. The decrease in the specific binding resulted from a lower number of binding sites and not from change in the dissociation constants. The decrease in the total receptors in the liver was more gradual and leveled off at 40 - 50% of the initial value. Our results suggest that a change in blood PRL or hGH level may lead to a new steady state in the number, occupancy and distribution of prolactin receptors.

Animals↗
PubMed1983

Human growth hormone but not ovine or bovine growth hormones exhibits galactopoietic prolactin-like activity in organ culture from bovine lactating mammary gland.

Explants from the mammary gland of 6 lactating cows were cultured in M-199 medium containing insulin (1.0 micrograms/ml) and hydrocortisone (0.5 micrograms/ml) and supplemented with bPRL (0.2-1.0 micrograms/ml) or oGH (0.1, 0.5 and 2.5 micrograms/ml) or bGH or hGH (0.2 and 1.0 micrograms/ml). It was found that neither oGH nor bGH significantly increased (P less than 0.05) the casein and fat synthesis and alpha-lactalbumin secretion above the control level. A significant and almost equal increase was, however, observed in the presence of bPRL and hGH, but was not influenced by the simultaneous addition of oGH to bPRL-containing medium (both at 0.5 micrograms/ml). Various hormonal treatments did not affect casein secretion and glucose uptake. High correlations (r = 0.89-0.97) were found between the mean values for casein and fat synthesis and alpha-lactalbumin secretion. Our results suggest that (a) the in vivo galactopoietic activity of bGH does not result from a direct effect of synthetic capacities on the mammary gland, and (b) hGH is a potent lactogen in the organ culture of bovine lactating mammary tissue.

Animals↗
PubMed1982

Hormonal control of casein synthesis in organ culture of the bovine lactating mammary gland.

Explants from lactating bovine mammary gland were cultured in vitro in serum-free medium though 1-9 d. casein synthesis was determined by [32P] incorporation into newly synthesized Ca rennin precipitable fraction. High correlation (r = 0.98) was found between incorporation of [32P] and [3H]amino acids in explants cultured under different hormonal regimes, thus indicating that post-translational phosphorylation is not a rate-limiting step in casein synthesis. Hormonal effects on casein synthesis were studied by supplementing the incubating medium with insulin (I), prolactin (PRL), cortisol (F), thyroxine (T4) and triiodothyronine (T3). It was found that both PRL and I were required absolutely for maximal synthesis and almost maximal effect was achieved with 50 ng/ml. The effect of F was less clear, but some increase was achieved at the 200-1000 ng/ml range. T4 and T3 did not affect casein synthesis at a range of 10(-11)-10(-7) M while a significant inhibition was observed at 2 X 10(-5) M. A time-course study of casein synthesis further substantiated the dominant role of PRL in maintenance or even elevation of the initial rate of casein synthesis in the explants, through the first 4 d of incubation.

Animals↗
PubMed1982

The effect of dietary raw and autoclaved soya-bean protein fractions on growth, pancreatic enlargement and pancreatic enzymes in rats.

1. Raw soya-bean meal (RS) was fractionated into soya-bean lyophilized extract (SLE), soya-bean lyophilized residue (SLR), acid-precipitated proteins (APP) and whey proteins. 2. Trypsin (EC 3. 4. 21. 4)and chymotrypsin (EC 3. 4. 21.1) inhibitors (TI) were soluble at pH 8 and remained soluble after the extract was acidified to pH 4.4 Except for whey, heating abolished, almost totally, their inhibiting activity. 3. Feeding SLE diet (high TI content) and APP diet (low TI content) resulted in growth depression below the RS level. Feeding the SLR diet resulted in an optimal growth. Feeding diets containing heated fractions improved the growth rate though not to the level observed with heated RS (HS) diet. 4. RS, SLE, APP and whey diets produced similar pancreatic enlargement which could be totally (RS, whey) or partially (SLE, APP) abolished by heating. 5. Feeding the RS diet reduced pancreatic amylase content. The factor responsible for this effect cofractionated with SLE and whey proteins. 6. Two groups of factors in the various diets were probably responsible for the elevation in pancreatic proteases. The first group were the heat-labile factors present in RS, SLE and whey whereas the second group resisted the heat treatment and were found in APP and SLR. 7. The results suggest that for optimal growth rate of rats, heat treatment should be given to the unfractionated soya-bean proteins rather than to the isolated fractions. The results further indicated that TI are not the only factors that can lead to pancreatic enlargement and changes in pancreatic enzymes composition.

Animals↗
PubMed1982

The kidney is the main site of interferon degradation.

Male Sprague Dawley rats (300 g) were infused by constant infusion into the jugular vein through 3-5 h with human leukocyte-derived interferon (HuIFN-alpha 10(6) units/h, 0.9 ml/h). Blood samples were collected every 20 min through the carotid. A steady-state level of interferon in serum (10(4) U/ml) was reached after 50 min of infusion, indicating fast removal of most of the infused material. Examination of interferon distribution in various tissues at the termination of the infusion revealed that over 85% of active interferon was found in the kidneys. No interferon was found in urine. Ligation of both kidneys resulted (after 2.5 h of infusion) in a ten-fold increase in interferon in serum and a minor increase in other tissues. No interferon was found in the kidneys. Simultaneous infusion of HuIFN-alpha, pepstatin and leupeptin (the well-known inhibitors of lysosomal proteinases) for 2.5 h had no effect on interferon concentration in serum and various tissues except in kidneys where a 5-fold increase was found. Mitochondrial-lysosomal fractions contained 55% of the total kidney interferon. Partial inhibition of proteolytic and HuIFN-alpha degrading activities at that fraction was also observed. These data, together with our previous findings of accumulation of injected interferon in the mitochondrial-lysosomal fraction of rats and monkeys' kidney cells, provide a further evidence for the main role of the kidney in HuIFN-alpha degradation.

Animals↗
PubMed1982

Involvement of the kidney in catabolism of human leukocyte interferon.

The metabolic fate of human leukocyte interferon (HuIFN-alpha) was studied after intravenous injection into rats and cynomolgus monkeys. At various intervals the animals were sacrificed and the HuIFN-alpha content determined in serum and various tissues. HuIFN-alpha quickly disappeared from the circulation and was found mainly in the kidneys, in which levels were at least 7- to 10-fold higher than in the liver, spleen, lungs, heart, brain and muscles. No interferon was detected in urine. Subcellular fractionation of kidney revealed that the mitochondrial-lysosomal fraction (15 000 g) had a high HuIFN-alpha content. It was also found that HuIFN-alpha was rapidly inactivated by two types of proteinases found in the lysosomal fractions of rat, monkey and human kidneys, with an optimal pH of 3 to 4. The inactivation was partially inhibited by either pepstatin or leupeptin. Inactivation was totally prevented by a mixture of both inhibitors. Since it is known that interferon is scantily excreted in urine, our findings suggest that the kidney serves as a main site for its degradation.

Animals↗
PubMed1981

Inhibition of porcine elastase II by chicken ovoinhibitor.

Interaction of porcine elastase II with native and modified chicken egg-white ovoinhibitor was studied by determining the residual activity of the partially inhibited enzyme and by direct measurement of the stoichiometry of interaction using affinity chromatography, electrophoresis and gel filtration. It was found that the chymotrypsin binding site that is not modified by mild oxidation with N-chlorosuccinimide (Shechter et al., Biochemistry, (1977) 16, 992-997) is capable of binding elastase II as well. The binding of chymotrypsin and elastase II is mutually exclusive and the affinity of chymotrypsin is stronger. Binding of 2 mol trypsin as 1 mol elastase I by ovoinhibitor does not interfere with the binding of elastase II. There is also an indication that the second binding site for chymotrypsin is capable of forming a complex with an additional molecule of elastase II, but the binding is so weak that it could be detected only by electrophoresis.

Amino Acids↗
PubMed1981

Urine and serum leucine aminopeptidase, N-acetyl-beta-glucosaminidase and gamma-glutamyl transpeptidase activities in diabetics with and without nephropathy.

Urinary and serum activities of leucine aminopeptidase (LAP), N-acetyl-beta-glucosaminidase (ABG) and gamma-glutamyl transpeptidase (GGTP) were determined in four groups: 1) control subjects with normal oral glucose tolerance test and normal renal function; 2) latent diabetics with elevated oral glucose tolerance test and diabetic parents; 3) overt diabetics; 4) diabetics with slight nephropathy, exemplified by elevated serum creatinine levels and decreased creatinine clearance. No clear differences were found in the urinary GGTP and serum enzyme activities. Urinary LAP activity was significantly increased in Groups 2, 3 and 4, and urinary ABG activity in Groups 3 and 4. However, since the rise in urinary LAP activity was less consistent than that in ABG, it seems that the appearance of the latter enzyme in urine most likely results from tubular damage and may serve as a sensitive indicator of early diabetic kidney disease.

Acetylglucosaminidase↗