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Query: HUMANGGP:034761 (insulin)
 211,843 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A plasma membrane-enriched fraction was prepared from homogenized rat pancreatic islets by a one-step sucrose gradient centrifugation. Using 125I-wheat germ agglutinin as a plasma membrane probe, a fraction was obtained at a sucrose density of about 1.10 that was enriched in 5'-nucleotidase, Mg2+-ATPase and alkaline phosphatase. The fraction contained little, if any, monoamino oxidase activity, insulin or DNA. Hydrolysis of 3-0-methyl-fluoresceinphosphate was stimulated by K+ (10mM) at a pH optimum of pH 8.2. Hydrolysis of ATP-gamma-32P in the presence of MgCl2 was of high specific activity and was optimum at pH 7.0 and 8.2. K+ did not affect ATP-hydrolysis. At pH 8.2, a small fraction of the total Mg2+-ATPase activity was inhibited by ouabain in the presence of Na+ and K+. Since K+-stimulated phosphatase activity does not correlate with Mg2+-ATPase, the two assay systems define separate enzymatic processes.
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PMID:Cation-dependent phosphatase activites in a rat pancreatic islet plasma membrane fraction prepared by one-step gradient centrifugation. 3 53

DNA synthetic activity was monitored in rat and human prostate using [125I] iododeoxyuridine ([125I]UdR). Fresh prostate tissue from 6-week-old rats showed higher incorporation of [125I]UdR than that from 12- or 26-week-old rats. During culture for up to 6 days in the absence of hormones, the incorporation of [125I]UdR fell to a low level for all three age groups. Stimulatory effects were seen when rat prostates were cultured in the presence of insulin (3 mug/ml) and testosterone (10(-7) mol/l), the incorporation on day 4 of culture being commensurate with that of fresh prostrate of the corresponding age. Thus the magnitude of the response was higher for the 6-week-old prostate than that for the other two age groups. A similar age-related pattern of androgen stimulation was observed in experiments in which immature and adult castrated rats were injected daily with testosterone and the freshly removed prostates were incubated with [125I]UdR. Although insulin, by itself, had a stimulatory effect on [125I]UdR incorporation in cultured prostate, the magnitude of the response did not differ in the 6- and 26-week-old prostate tissue. Maximal stimulation was obtained using 25 mug insulin/ml. Tissue from a benigh prostatic hyperplasia was also responsive to insulin in culture but it differed from rat prostate in that increased proliferative activity occurred even in the absence of hormone stimulation. This spontaneous surge in activity during culture tended to mask the stimulatory effects of insulin and testosterone at concentrations of 3 mug/ml and 10(-7) mol/l respectively.
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PMID:Uptake of (125I) iododeoxyuridine in cultured rat and human prostate: effects of insulin and testosterone. 5 51

When Vero cells, a line derived from and African Green Monkey kidney, are grown under conditions where the saturation density is limited by serum, they deplete the growth medium of a factor necessary for cell division. The factor is a component of serum. When Vero cells are plated at low density (2 X 10(4)/cm2) in this depleted growth medium (after dialysis against serum-free Dulbecco's Modified Eagle's Medium) they initiate an unbalanced program of growth. Protein synthesis proceeds at the same rate as parallel cells in fresh serum, and and the cells accumulate protein as a function of time. DNA synthesis is also initiated in these cells, and the amount of DNA per cell increases for the next four days plating. However the cells quickly stop dividing. Measurements of DNA per cell using microspectrofluorometry show that the cells are accumulating in the late S and G2 period during this time. Thus we conclude that these cells cannot pass through a transition point in G2. When fresh serum is added to cells after three days in depleted growth medium, they divide before they begin to synthesize DNA. This further confirms that they are in late S and G2. Cell division is promoted in Vero cells in depleted growth medium by bovine fetuin, and to a lesser extent by bovine albumin. Cell division is not promoted by insulin, hydrocortisone, dexamethasone, linolenic acid, calcium, and typsin inhibitor form ovomucoid. From these data we conclude that transit through G2 requires the prescence of an extracellular factor.
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PMID:A serum factor requirement for the passage of cultured Vero cells through G2. 6 56

DNA sequences corresponding to specific genes may be prepared by chemical synthesis, isolation of naturally occurring DNA, or reverse transcription. Such DNA may then be inserted into vectors such as plasmids or bacteriophages which carry the DNA into bacterial cells. Although significant differences exist in the basic molecular biology of eucaryotic and procaryotic organisms, these differences do not constitute absolute barriers to the expression of eucaryotic genes in bacteria. Several eucaryotic proteins, including insulin, growth hormone, ovalbumin, dihydrofolate reductase and somatostatin have been produced in bacteria. The use of chimeric microorganisms harboring recombinant DNA offers a completely new approach to the production of biologically useful polypeptides.
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PMID:Use of recombinant DNA technology for the production of polypeptides. 9 11

T-cell DNA synthesis and T-helper cell function in response to isolated insulin chains and naturally occurring insulin variants was assessed in insulin immune guinea pigs. Two distinct antigenic determinants, recognized by T cells, were defined. One localized in the B chain and the other one constituted by amino acids A8, A9, and A10 of the insulin A-chain loop. Recognition of the B-chain determinant is under the control of Ir genes linked to the strain 13 major histocompatibility complex. This was shown by studying the response to isolated insulin B chain in F1(2 x 13) guinea pigs, as well as serologically defined backcrosses and outbred animals. Insulin recognition through the A-chain loop determinant is specific for strain 2 guinea pigs. These animals recognize this region of the molecule even when displaying different amino acid sequences. The strain differences observed in those antigenic sites eliciting T-cell recognition was not found at an antibody level. No differences could be detected in the ability of the different insulin variants to inhibit the binding of 125I-labeled pork insulin to strain 2 guinea pig anti-pork insulin or to strain 13 guinea pig anti-pork insulin.
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PMID:Immune response gene control of determinant selection. I. Intramolecular mapping of the immunogenic sites on insulin recognized by guinea pig T and B cells. 9 87

The mammary glands of euthyroid female C3H/HeN mice undergo a series of morphological changes during development. In glands from immature animals, the epithelial component consists of a sparse ductal system with few branches which fills about one fourth of the fat pad. In the adult virgin gland, the epithelial component fills the fat pad with a highly branched ductal system and a few alveoli. In contrast, glands from adult animals maintained in a hypothyroid state by ingestion of thiouracil since weaning retain the primitive ductal appearance while filling the fat pad. The glands from animals made hyperthroid by adding 2 micrograms T4/ml drinking water have extensive lobulo-alveolar development. Glands from animals made hypothyroid during 7 weeks of involution after lactation have the same degree of deveopment as the euthyroid controls. When explants of tissue from adult hypothyroid virgin animals are cultured in serum-free medium containing insulin, hydrocortisone, and PRL, the specific milk protein, alpha-lactalbumin, is induced. The level of alpha-lactalbumin, measured as lactose synthetase activity, found per ng epithelial DNA is the same as that found in explants from glands of euthyroid virgins. These results suggest that thyroid hormones, in concert with PRL, play an important role in the regulation of development of the mouse mammary gland. Decreased levels of thyroid hormones in the serum result in retarded growth of the ductal system and little or no alveolar development. However, the resulting epithelial component of glands from hypothyroid mice is fully capable of differentiating in vitro when exposed to the proper hormonal environment.
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PMID:Lobulo-alveolar development of mouse mammary glands is regulated by thyroid hormones. 10 79

Dimethylbenz(alpha)anthracene (DMBA)-induced and transplanted rat mammary tumours (2 lines) were examined for oestrogen receptor activity, and for sensitivity to hormones in vivo (by ovariectomy) and in vitro (by tissue culture). In vivo, the growth of all tumours induced by the administration of DMBA in random-bred Sprague-Dawley rats was found to be dependent on the ovary, whilst in all transplanted tumours (12 TG-3 and six TG-5 lines), maintained in an inbred strain of Sprague-dawley rats, growth was found to be independent of the ovary. In vitro, the capacity for DNA synthesis in DMBA-induced tumours was better maintained after 24 h when insulin (10 microgram/ml) and corticosterone (5 microgram/ml) or insulin, corticosterone and prolactin (each 5 microgram/ml) were present in the medium (five out of 12 and eight out of 11 tumours respectively); no effect of hormones in the media was detected after 48 h. In the transplanted tumours, no effect of hormones on DNA synthesis was detected after either 24 or 48 h of culture. Synthesis of lecithin was not detectably influenced by the presence of hormones in either DMBA-induced or transplanted tumours. Oestrogen receptor concentrations were, on average, significantly higher in the DMBA-induced tumours than in either line of transplanted tumour. For 22 DMBA-induced tumours and 15 transplanted tumours, the effect of hormones in vitro ('response') was directly correlated with receptor concentration at time 0 (Spearman's rho = +0.59) and inversely correlated with the rate of DNA synthesis ('basal') at time 0 (Spearman's rho = -0.62). No single parameter or pair of parameters permitted accurate distinction between the tumour types.
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PMID:Synthesis of DNA and lecithin in tissue culture and oestrogen receptor activity in rat mammary tumours dependent on and independent of the ovary. 11 Sep 1

Epithelial cells in explants from the mammary glands of euthyroid mature virgin mice are proliferatively dormant. They must undergo DNA synthesis and traverse the cell cycle in vitro before they are able to differentiate fully in response to insulin, hydrocortisone, and prolactin, and synthesize enzymatically active alpha-lactalbumin (measured as lactose synthetase activity). In contrast, glands from hyperthyroid mature virgin mice do not require DNA synthesis in vitro to differentiate. Explants from the euthyroid virgin tissue overcome their dependence on DNA synthesis when 10(-9) M 3,5,3'-triiodo-L-thyronine is added directly to the cultures in addition to the other three hormones. Explants from involuted mammary glands from euthyroid primiparous mice do not require DNA synthesis in vitro to make the milk protein even though they, like explants from mature euthyroid virgin tissue, are proliferatively dormant and do not contain detectable lactose synthetase activity in vivo. Glands from primiparous animals made mildly hypothyroid by ingestion of 0.1% thiouracil in drinking water during 7 wk of involution remain morphologically indistinguishable from glands of their euthyroid counterparts. However, explants from the glands of these hypothyroid animals revert to a state of dependence on DNA synthesis to differentiate functionally. These observations suggest that the dependence on DNA synthesis and cell cycle traversal for hormonal induction of lactose synthetase activity in the mouse mammary gland is controlled by thyroid hormones.
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PMID:Lactose synthetase activity in mouse mammary glands is controlled by thyroid hormones. 11 14

Skeletal muscle can undergo rapid growth in response to a sudden increase in work load. For example, the rat soleus muscle increases in weight by 40% within six days after the tendon of the synergistic gastrocnemius is sectioned. Such growth of the overworked muscle involves an enlargement of muscle fibers and occasional longitudinal splitting. Hypertrophy leads to greater maximal tension development, although decreased contraction time and reduced contractility have also been reported. Unlike normal developmental growth, work-induced hypertrophy can be induced in hypophysectomized or diabetic animals. This process thus appears independent of growth hormone and insulin as well as testosterone and thyroid hormones. Hypertrophy of the soleus can also be induced in fasting animals, in which there is a generalized muscle wasting. Thus muscular activity takes precedence over endocrine influences on muscle size. The increase in muscle weight reflects an increase in protein, especially sarcoplasmic protein, and results from greater protein synthesis and reduced protein breakdown. Within several hours after operation, the hypertrophying soleus shows more rapid uptake of certain amino acids and synthesis of phosphatidyl-inositol. By 8 hours, protein synthesis is enhanced. RNA synthesis also increases, and hypertrophy can be prevented with actinomycin D. Nuclear DNA synthesis also increases on the second day after operation and leads to a greater DNA content. The significance of the increased RNA and DNA synthesis is not clear, since most of it occurs in interstitial and satellite cells. The proliferation of the non-muscle cells seems linked to the growth of the muscle fibers; in addition, factors causing muscle atrophy (e.g. denervation) decrease DNA synthesis by such cells. In order to define more precisely the early events in hypertrophy, the effects of contractile activity were studied in rat muscles in vitro. Electrical stimulation enhanced active transport of certain amino acids within an hour, and the magnitude of this effect depended on the amount of contractile activity. Stimulation or passive stretch of the soleus or diaphragm also retarded protein degradation. Presumably these effects of mechanical activity contribute to the changes occuring during hypertrophy in vivo. However, under the same conditions, or even after more prolonged stimulation, no change in rates of protein synthesis was detected. These findings with passive tension in vitro are particularly interesting, since passive stretch has been reported to retard atrophy or to induce hypertrophy of denervated muscle in vivo. It is suggested that increased tension development (either passive or active) is the critical event in initiating compensatory growth.
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PMID:Mechanism of work-induced hypertrophy of skeletal muscle. 12 81

1. Kidney weight and content of protein, RNA and DNA were measured in rats with streptozotocin diabetes of varying duration. 2. Diabetic rats had larger kidneys than control rats: after 3 days of diabetes the weight increase was 15 per cent and after 42 days of diabetes it was 90 per cent. The protein content rose in parallel to the weight. 3 RNA content was already increased after 36 h of glycosuria, whereas DNA content was unchanged for the first 3 days of diabetes, and increased thereafter. The protein/DNA ratio increased rapidly during the first 3 days but remained constant thereafter. 4. Insulin treatment decreased the renal weight gain by about 67 per cent during the first 8 days of diabetes, but did not prevent the increase in DNA. When insulin was started after 25 days of diabetes there was only a slight regression of kidney growth.
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PMID:Renal hypertrophy in streptozotocin-diabetic rats. 13 97


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