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1.
IEEE Trans Neural Netw ; 19(2): 333-45, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18269963

ABSTRACT

In this paper, we analyze a model of recurrent kernel associative memory (RKAM) recently proposed by Garcia and Moreno. We show that this model consists in a kernelization of the recurrent correlation associative memory (RCAM) of Chiueh and Goodman. In particular, using an exponential kernel, we obtain a generalization of the well-known exponential correlation associative memory (ECAM), while using a polynomial kernel, we obtain a generalization of higher order Hopfield networks with Hebbian weights. We show that the RKAM can outperform the aforementioned associative memory models, becoming equivalent to them when a dominance condition is fulfilled by the kernel matrix. To ascertain the dominance condition, we propose a statistical measure which can be easily computed from the probability distribution of the interpattern Hamming distance or directly estimated from the memory vectors. The RKAM can be used below saturation to realize associative memories with reduced dynamic range with respect to the ECAM and with reduced number of synaptic coefficients with respect to higher order Hopfield networks.


Subject(s)
Association Learning/physiology , Memory/physiology , Neural Networks, Computer , Computer Simulation , Humans , Models, Neurological , Recurrence
2.
J Mol Endocrinol ; 34(2): 367-76, 2005 Apr.
Article in English | MEDLINE | ID: mdl-15821103

ABSTRACT

The elucidation of mechanisms regulating the regeneration and survival of pancreatic beta cells has fundamental implications in the cell therapy of type 1 diabetes. The present study had the following three aims: 1. to investigate whether pancreatic ductal epithelial cells can be induced to differentiate into insulin-producing cells by exposing them to hepatocyte growth factor (HGF); 2. to characterize some of the molecular events leading to their differentiation toward a beta-cell-like phenotype; 3. to evaluate the susceptibility of newly differentiated insulin-secreting cells to cytokine-induced apoptosis, a mechanism of beta-cell destruction occurring in type 1 diabetes. We demonstrated that HGF-treated rat pancreatic ductal cell line (ARIP) cells acquired the capability to transcribe the insulin gene and translate its counterpart protein. HGF-treated cells also exhibited a glucose-dependent capability to secrete insulin into the cultured medium. Expression analysis of some of the genes regulating pancreatic beta-cell differentiation revealed a time-dependent transcription of neurogenin-3 and Neuro-D in response to HGF. Finally, we determined the susceptibility to proinflammatory cytokine (PTh1)-induced apoptosis by incubating HGF-treated and untreated ARIP cells with a cocktail of interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). Such treatment induced apoptotic death, as determined by the TUNEL technique, in about 40% of HGF-treated, insulin-secreting ARIP cells, while untreated ARIP cells were resistant to PTh1-induced apoptosis. In conclusion, we showed that HGF promotes the differentiation of ARIP cells into pancreatic beta-cell-like cells, and that the differentiation toward an insulin-secreting phenotype is associated with the appearance of susceptibility to cytokine-induced apoptosis.


Subject(s)
Apoptosis/physiology , Cytokines/pharmacology , Epithelial Cells/drug effects , Hepatocyte Growth Factor/pharmacology , Insulin/metabolism , Pancreatic Ducts/cytology , Animals , Basic Helix-Loop-Helix Transcription Factors , Cell Differentiation/physiology , Cells, Cultured , Cytokines/metabolism , Epithelial Cells/cytology , Epithelial Cells/metabolism , Gene Expression Regulation , Glucose/metabolism , Humans , Insulin/genetics , Insulin Secretion , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Islets of Langerhans/cytology , Islets of Langerhans/physiology , Models, Biological , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Phenotype , Rats , Receptor, Notch3 , Receptors, Cell Surface/genetics , Receptors, Cell Surface/metabolism , Receptors, Notch , Transcription Factors/genetics , Transcription Factors/metabolism
3.
Horm Metab Res ; 36(11-12): 804-10, 2004.
Article in English | MEDLINE | ID: mdl-15655712

ABSTRACT

Glucagon-like peptide-1 (GLP-1), a peptide hormone produce by intestinal cells, has recently been shown to be capable of modulating islet cell mass. Administration of GLP-1 to rodent models of type 2 diabetes ameliorates insulin secretion, induces the replication of islet cells, and promotes islet-cell neogenesis from pancreatic ductal cells susceptible to transdifferentiate in insulin-producing cells. In addition, an anti-apoptotic effect of GLP-1 has been described in hyperglycemic animal models, using freshly isolated human islets or cultured beta cell lines exposed to various pro-apoptotic stimuli. The aim of this article is to review those reports that have emphasized the role of GLP-1 as a regulator of islet cell mass.


Subject(s)
Apoptosis/physiology , Glucagon/physiology , Islets of Langerhans/physiology , Peptide Fragments/physiology , Protein Precursors/physiology , Animals , Cell Differentiation/physiology , Glucagon-Like Peptide 1 , Glucose Transporter Type 2 , Humans , Insulin/genetics , Insulin/metabolism , Islets of Langerhans/cytology , Monosaccharide Transport Proteins/genetics , Monosaccharide Transport Proteins/metabolism , Rats
4.
IEEE Trans Neural Netw ; 14(3): 703-7, 2003.
Article in English | MEDLINE | ID: mdl-18238051

ABSTRACT

We present a neural associative memory storing gray-scale images. The proposed approach is based on a suitable decomposition of the gray-scale image into gray-coded binary images, stored in brain-state-in-a-box-type binary neural networks. Both learning and recall can be implemented by parallel computation, with time saving. The learning algorithm, used to store the binary images, guarantees asymptotic stability of the stored patterns, low computational cost, and control of the weights precision. Some design examples and computer simulations are presented to show the effectiveness of the proposed method.

5.
J Mol Endocrinol ; 29(3): 347-60, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12459036

ABSTRACT

The intestinal hormone glucagon-like peptide-1 (GLP-1) has been shown to promote an increase in pancreatic beta-cell mass via proliferation of islet cells and differentiation of non-insulin-secreting cells. In this study, we have characterized some of the events that lead to the differentiation of pancreatic ductal cells in response to treatment with human GLP-1. Rat pancreatic ductal (ARIP) cells were cultured in the presence of GLP-1 and analyzed for cell counting, cell cycle distribution, expression of cyclin-dependent-kinase (Cdk) inhibitors, transcription of beta-cell-specific genes, loss of ductal-like phenotype and acquisition of beta-cell-like gene expression profile. Exposure of ARIP cells to 10 nM GLP-1 induced a significant reduction in the cell replication rate and a significant decrease in the percentage of cells in S phase of the cell cycle. This was associated with an increase in the number of cells in G0-G1 phase and a reduction of cells in G2-M phase. Western blot analysis for the Cdk inhibitors, kinase inhibitor protein 1 (p27(Kip1)) and Cdk-interacting protein 1 (p21(Cip1)), demonstrated a significant increase in p27(Kip1) and p21(Cip1) levels within the first 24 h from the beginning of GLP-1 treatment. As cells slowed down their proliferation rate, GLP-1 also induced a time-dependent expression of various beta-cell-specific mRNAs. The glucose transporter GLUT-2 was the first of those factors to be expressed (24 h treatment), followed by insulin (44 h) and finally by the enzyme glucokinase (56 h). In addition, immunocytochemistry analysis showed that GLP-1 induced a time-dependent down-regulation of the ductal marker cytokeratin-20 (CK-20) and a time-dependent induction of insulin expression. Finally, GLP-1 promoted a glucose-dependent secretion of insulin, as demonstrated by HPLC and RIA analyses of the cell culture medium. The present study has demonstrated that GLP-1 induces a cell cycle re-distribution with a decrease in cell proliferation rate prior to promoting the differentiation of cells towards an endocrine-like phenotype.


Subject(s)
Cell Cycle/drug effects , Cell Differentiation/drug effects , Glucagon/pharmacology , Pancreatic Ducts/cytology , Pancreatic Ducts/drug effects , Peptide Fragments/pharmacology , Protein Precursors/pharmacology , Animals , Cell Cycle Proteins/metabolism , Cells, Cultured , Cyclin-Dependent Kinase Inhibitor p21 , Cyclin-Dependent Kinase Inhibitor p27 , Cyclins/metabolism , Flow Cytometry , Gene Expression Regulation/drug effects , Glucagon-Like Peptide 1 , Glucokinase/metabolism , Glucose/pharmacology , Glucose Transporter Type 2 , Humans , Immunohistochemistry , Insulin/analysis , Insulin/metabolism , Insulin Secretion , Monosaccharide Transport Proteins/metabolism , Rats , Tumor Suppressor Proteins/metabolism
6.
J Mol Endocrinol ; 27(2): 133-44, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11564599

ABSTRACT

The Arg64 beta(3)-adrenergic receptor (beta(3)AR) variant is associated with an earlier age of onset of diabetes and lower levels of insulin secretion in humans. The aims of this study were to investigate whether beta(3)AR is expressed by islet cells, if receptor binding affects insulin secretion and, finally, if the beta(3)AR Arg64 variant induces abnormal insulin secretory activity. Human pancreas extracts were subjected to RT-PCR, Western blotting and immunostaining analyses. DNA sequencing and Western blotting demonstrated that the beta(3)AR gene is transcribed and translated in the human pancreas; immunostaining showed that it is expressed by the islets of Langerhans. Cultured rat beta-cells responded to human beta(3)AR agonists in a dose- and time-dependent manner. Transfection of cultured rat beta-cells with the wild-type human beta(3)AR produced an increased baseline and ligand-dependent insulin secretion compared with parental cells. On the other hand, cells transfected with the Arg64 variant of the beta(3)AR secreted less insulin, both spontaneously and after exposure to human beta(3)AR agonists. Furthermore, while transfection with the wild-type beta(3)AR preserved the glucose-dependent secretion of insulin, expression of the variant receptor rendered the host cells significantly less responsive to glucose. In summary, cells express the beta(3)AR, and its activation contributes to the regulation of insulin secretion. These findings may help explain the low levels of insulin secretion in response to an i.v. glucose tolerance test observed in humans carrying the Arg64 polymorphism.


Subject(s)
Genetic Variation , Insulin/metabolism , Islets of Langerhans/metabolism , Receptors, Adrenergic, beta-3/genetics , Adrenergic beta-3 Receptor Agonists , Adrenergic beta-Agonists/metabolism , Adrenergic beta-Agonists/pharmacology , Animals , Arginine/genetics , Base Sequence , Cell Line , DNA Primers/genetics , Dioxoles/pharmacology , Ethanolamines/metabolism , Ethanolamines/pharmacology , Gene Expression , Humans , Insulin Secretion , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Receptors, Adrenergic, beta-3/metabolism , Transfection
7.
J Endocrinol Invest ; 24(4): 274-87, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11383915

ABSTRACT

Type 2 diabetes is characterized by two fundamental biological defects: a reduced glucose-dependent insulin secretion and an increased resistance to the action of insulin at the level of various target tissues. While the use of agents to improve the insulin secretory activity of the islets of Langerhans has witnessed the flourishing of several new drugs over the years, a much greater difficulty has been experienced in the search for insulin-sensitizing drugs. The aim of this article is to critically review this topic, and to emphasize the importance of providing alternative strategies for the management of Type 2 diabetes.


Subject(s)
Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/therapeutic use , Insulin/physiology , Thiazoles/therapeutic use , Animals , Humans , Minerals/therapeutic use
8.
Diabetes ; 50(4): 785-96, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11289043

ABSTRACT

Glucagon-like peptide-1 (GLP-1) is an incretin hormone capable of restoring normal glucose tolerance in aging glucose-intolerant Wistar rats. Whether the antidiabetic properties of GLP-1 are exclusively due to its insulin secretory activity remains to be determined. A GLP-1-dependent differentiation of pancreatic precursor cells into mature beta-cells has recently been proposed. The aim of this study was to investigate whether pancreatic ductal epithelial cells could be differentiated into insulin-secreting cells by exposing them to GLP-1. Rat (ARIP) and human (PANC-1) cell lines, both derived from the pancreatic ductal epithelium, were used to test this hypothesis. A major difference distinguishes these two cell lines: whereas ARIP cells spontaneously express the beta-cell differentiation factor islet duodenal homeobox-1 (IDX-1), PANC-1 cells are characteristically IDX-1 negative. GLP-1 induced the differentiation of ARIP cells into insulin-synthesizing cells, although it did not affect the phenotype of PANC-1 cells, as determined by fluorescence-activated cell sorting (FACS) analysis. Differentiation of ARIP cells by exposure to human GLP-1 occurs in a time- and dose-dependent manner, and this is associated with an increase in IDX-1 and insulin mRNA levels. Secretion of insulin was also induced in a parallel manner, and it was regulated by the concentration of glucose in the culture medium. Interestingly, PANC-1 cells, when stably transfected with human IDX-1, gained responsiveness to GLP-1 and were able to differentiate into beta-cells, as determined by FACS analysis, insulin gene expression, intracellular insulin content, and insulin accumulation in the culture medium. Finally, we demonstrated that the receptor for GLP-1 is constitutively expressed by ARIP and PANC-1 cells and that the mRNA level for this transcript was increased by cellular transfection with human IDX-1. In summary, our study provides evidence that GLP-1 is a differentiation factor for pancreatic ductal cells and that its effect requires the expression of IDX-1.


Subject(s)
Glucagon/pharmacology , Homeodomain Proteins , Pancreatic Ducts/cytology , Pancreatic Ducts/metabolism , Peptide Fragments/pharmacology , Protein Precursors/pharmacology , Trans-Activators/metabolism , Actins/genetics , Animals , Cell Differentiation/drug effects , Cell Line , Culture Media/metabolism , Flow Cytometry , Fluorescent Antibody Technique , Glucagon-Like Peptide 1 , Humans , Immunohistochemistry , Insulin/metabolism , RNA, Messenger/metabolism , Rats , Trans-Activators/genetics
9.
Eur J Endocrinol ; 143(6): 717-25, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11124853

ABSTRACT

Glucagon-like peptide-1 (GLP-1) is a gut hormone synthesized by post-translational processing in intestinal L-cells, and it is released in response to food ingestion. GLP-1 stimulates insulin secretion during hyperglycemia, suppresses glucagon secretion, stimulates (pro)-insulin biosynthesis and decreases the rate of gastric emptying and acid secretion. GLP-1 has also been shown to have a pro-satiety effect. In addition, it has been demonstrated that a long-term infusion with GLP-1, or exendin-4, a long-acting analog of human GLP-1, increases beta-cell mass in rats. In conclusion, GLP-1 appears to regulate plasma glucose levels via various and independent mechanisms. GLP-1 is an excellent candidate option for the treatment of patients with type 2 diabetes mellitus.


Subject(s)
Insulin-Like Growth Factor I/physiology , Islets of Langerhans/physiology , Animals , Gene Expression Regulation/physiology , Glucagon/metabolism , Humans , Hyperglycemia/physiopathology , Insulin/metabolism , Insulin Secretion , Insulin-Like Growth Factor I/genetics , Islets of Langerhans/physiopathology
10.
Endocrinology ; 141(12): 4600-5, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11108273

ABSTRACT

Glucose homeostasis in mammals is maintained by insulin secretion from the beta-cells of the islets of Langerhans. Type 2 diabetes results either from primary beta-cell failure alone and/or a failure to secrete enough insulin to overcome insulin resistance. Here, we show that continuous infusion of glucagon-like peptide-1 (7-36) (GLP-1; an insulinotropic agent), to young and old animals, had effects on the beta-cell of the pancreas other than simply on the insulin secretory apparatus. Our previous studies on a rodent model of glucose intolerance, the aging Wistar rat, show that a plateau in islet size, insulin content, and beta-cell mass is reached at 13 months, despite a continuing increase in body weight. Continuous sc infusion of GLP-1 (1.5 pM/kg x min), over 5 days, resulted in normal glucose tolerance. Our current results in both young and old rats demonstrate that treatment caused an up-regulation of pancreatic-duodenum homeobox-1 (PDX-1) expression in islets and total pancreas, induced pancreatic cell proliferation, and beta-cell neogenesis. The effects on levels of PDX-1 messenger RNA were abrogated by simultaneous infusion of Exendin (9-39), a specific antagonist of GLP-1. PDX-1 protein levels increased 4-fold in whole pancreata and 6-fold in islets in response to treatment. Beta-cell mass increased to 7.2 +/- 0.58 from 4.88 +/- 0.38 mg, treated vs. control, respectively, P < 0.02. Total pancreatic insulin content also increased from 0.55 +/- 0.02 to 1.32 +/- 0.11 microg/mg total pancreatic protein. Therefore, GLP-1 would seem to be a unique therapy that can stimulate pancreatic cell proliferation and beta-cell differentiation in the pancreas of rodents.


Subject(s)
Aging , Cell Division/drug effects , Glucagon/pharmacology , Glucose Intolerance , Islets of Langerhans/drug effects , Peptide Fragments/pharmacology , Protein Precursors/pharmacology , Trans-Activators/genetics , Animals , Blood Glucose/analysis , Gene Expression Regulation/drug effects , Glucagon-Like Peptide 1 , Homeodomain Proteins/genetics , Insulin/blood , Islets of Langerhans/cytology , Islets of Langerhans/metabolism , Pancreas/cytology , Pancreas/drug effects , Pancreas/metabolism , Rats , Rats, Wistar
11.
Trends Endocrinol Metab ; 11(6): 218-23, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10878751

ABSTRACT

The onset of type 2 diabetes is characterized by two determining factors: the insufficient ability to secrete insulin and/or the resistance to its biological action. Although in a very small proportion of individuals, one of those two metabolic abnormalities is the leading cause of diabetes, in most subjects, the coexistence of both appears to be necessary for the clinical manifestation of diabetes. Current biomedical research continues to clarify the relative contributions of these defects to the pathogenesis of type 2 diabetes, and novel pharmacological agents are specifically designed to correct either the impaired insulin secretory activity or the resistance to the action of insulin. The aim of this article is to provide a critical review of new sulfonylurea and non-sulfonylurea drugs that have been recently introduced for the treatment of diabetes, as well as drugs that are still under investigation and are likely to be made available in the near future.


Subject(s)
Hypoglycemic Agents/pharmacology , Insulin/metabolism , Sulfonylurea Compounds/pharmacology , Humans , Insulin Secretion , Stimulation, Chemical
12.
J Clin Invest ; 105(7): 955-65, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10749575

ABSTRACT

The pancreatic processing enzymes, PC1 and PC2, convert proinsulin to insulin and convert proglucagon to glucagon and glucagon-like peptide 1 (GLP-1). We examined the effect of streptozotocin (STZ) treatment on the regulation of these enzymes and the production of insulin, glucagon, and GLP-1 in the rat. Pancreatic PC1 and PC2 mRNA increased >2-fold and >4-fold, respectively, in rats receiving intraperitoneal STZ (50 mg/kg) daily for 5 days. Immunocytochemistry revealed that, although pancreatic islet cells in the STZ-treated rats were sparse and atrophic PC1, PC2, glucagon, and GLP-1 immunoreactivity increased dramatically in the remaining islet cells. Heightened PC1 and PC2 expression was seen in cells expressing glucagon but not in insulin-expressing cells. Furthermore, in STZ-treated rats, bioactive GLP-1(7-36 amide) accumulated in pancreatic extracts and serum 3- and 2.5-fold, respectively, over control animals. This treatment also caused a 2-fold increase in the ratio of amidated forms of GLP-1 immunoreactivity to total glucagon immunoreactivity in the pancreas but did not affect the ratio of proinsulin to insulin. We conclude that hyperglycemic rats have an increased expression of prohormone converting enzymes in islet alpha cells, leading to an increase in amidated GLP-1, which can then exert an insulinotropic effect on the remaining beta cells.


Subject(s)
Aspartic Acid Endopeptidases/metabolism , Diabetes Mellitus, Experimental/metabolism , Glucagon/metabolism , Pancreas/enzymology , Peptide Fragments/metabolism , Protein Precursors/metabolism , Subtilisins/metabolism , Animals , Aspartic Acid Endopeptidases/genetics , Body Weight/drug effects , Diabetes Mellitus, Experimental/pathology , Diabetes Mellitus, Experimental/physiopathology , Glucagon-Like Peptide 1 , Immunohistochemistry/methods , Insulin/metabolism , Islets of Langerhans , Male , Pancreas/pathology , Proinsulin/metabolism , Proprotein Convertase 2 , Proprotein Convertases , RNA, Messenger , Rats , Rats, Wistar , Streptozocin , Subtilisins/genetics
13.
Diabetes Technol Ther ; 1(3): 297-305, 1999.
Article in English | MEDLINE | ID: mdl-11475276

ABSTRACT

In humans as well as in other animal species, the ingestion of food provides the fundamental source of energy for various cellular activities. The intake of food and the ability of controlling the plasma levels of substrates for energy production involve complex mechanisms that ensure a constantly adequate supply of metabolites both in the fasting and in the fed state. A number of hormonal peptides released from the gastrointestinal (GI) tract in response to the ingestion of food have been shown to play a critical role in the postprandial control of glucose homeostasis. They are known to act through three main mechanisms of action. These include; (1) stimulation of insulin secretion of pancreatic islet (beta) cells; (2) inhibition of hepatic gluconeogenesis by suppression of glucagon secretion; and (3) inhibition of GI motility. While for some of these hormones all three mechanisms of action are utilized under physiological conditions, others preferentially use one or a combination of two mechanisms for lowering postprandial hyperglycemia. Although the term glucoincretins (or incretins, or insulinotropic hormones) etymologically only describes factors capable of inducing insulin secretion, it is more frequently used to identify a larger class of peptides that, rather than manifesting a specific mechanism of action (i.e., insulin secretion), share the ability of controlling glucose excursion in the fed state (with or without a direct insulinotropic effect). The latter more inclusive meaning, incretins, is used in this article. This review summarizes recent advances on synthesis, secretion, blood plasma patterns, and metabolism of some of the major GI regulatory peptides acting in the postprandial state.


Subject(s)
Blood Glucose/metabolism , Gastrointestinal Hormones/physiology , Glucose/metabolism , Animals , Energy Metabolism , Gastrointestinal Motility , Glucagon/metabolism , Glucagon-Like Peptide 1 , Glucagon-Like Peptides , Gluconeogenesis , Homeostasis , Humans , Insulin/metabolism , Insulin Secretion , Islets of Langerhans/metabolism , Liver/metabolism , Peptide Fragments/physiology
14.
Mol Cell Endocrinol ; 157(1-2): 171-80, 1999 Nov 25.
Article in English | MEDLINE | ID: mdl-10619408

ABSTRACT

Oxidative damage to DNA has been documented in cells isolated from subjects with diabetes. Herein, we evaluate the mechanism(s) that regulate the expression of the DNA repair enzyme XPD. CHO cells transfected with the human insulin receptor (CHO/HIRc) showed a threefold increase in the level of XPD mRNA when compared to control CHO/neo cells (P < 0.01). The addition of insulin to serum-starved cells led to an increase in XPD mRNA levels in both CHO/neo and CHO/HIRc cells, in a time and dose dependent fashion. Insulin acted primarily by inducing XPD transcription. Moreover, inhibition of protein synthesis by cyclohexamide induced a marked degradation of XPD mRNA levels in insulin treated cells. Site-directed mutagenesis of the tyrosine-kinase domain of the insulin receptor abolished the increase in XPD mRNA resulting from the transfection with wild type insulin receptors (P < 0.001). Western blot analysis of cell extracts from CHO/neo and CHO/HIRc cells revealed an increase in XPD counterpart protein was also induced by transfecting cells with the human insulin receptor. Evaluation of DNA damage by means of internucleosomal fragmentation showed a dramatic decrease in DNA fragmentation in CHO cells transfected with wild-type insulin receptor compared to control CHO/neo cells. DNA fragmentation was further decreased by the addition of insulin in the culture medium. In summary, our data indicates that activation of the insulin receptor plays an important role in the cellular response leading to repair of damaged DNA.


Subject(s)
DNA Helicases , DNA Ligases/biosynthesis , DNA-Binding Proteins , Protein Biosynthesis , Receptor Protein-Tyrosine Kinases/pharmacology , Receptor Protein-Tyrosine Kinases/physiology , Transcription Factors , Animals , CHO Cells , Catalytic Domain/genetics , Cricetinae , Culture Media, Serum-Free , DNA Fragmentation/drug effects , DNA Ligases/drug effects , DNA Repair/genetics , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Insulin/metabolism , Insulin/pharmacology , Mutation , Proteins/drug effects , Proteins/genetics , RNA, Messenger/analysis , RNA, Messenger/drug effects , Receptor, Insulin/metabolism , Signal Transduction/physiology , Xeroderma Pigmentosum Group D Protein
15.
Diabetes Metab Rev ; 14(3): 207-25, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9816470

ABSTRACT

Diabetes mellitus is the most common endocrine disease, accounting for over 200 million people affected worldwide. It is characterized by a lack of insulin secretion and/or increased cellular resistance to insulin, resulting in hyperglycemia and other metabolic disturbances. People with diabetes suffer from increased morbidity and premature mortality related to cardiovascular, microvascular and neuropathic complications. The Diabetes Control and Complication Trial (DCCT) has convincingly demonstrated the relationship of hyperglycemia to the development and progression of complications and showed that improved glycemic control reduced these complications. Although the DCCT exclusively studied patients with Type 1 diabetes, there is ample evidence to support the belief that the same relationship between metabolic control and clinical outcome exists in patients with Type 2 diabetes. Therefore, a major effort should be made to develop and implement more effective treatment regimes. This article reviews those novel drugs that have been recently introduced for the management of Type 2 diabetes, or that have reached an advanced level of study and will soon be proposed for preliminary clinical trials. They include: (i) compounds that promote the synthesis/secretion of insulin by the beta-cell; (ii) inhibitors of the alpha-glucosidase activity of the small intestine; (iii) substances that enhance the action of insulin at the level of the target tissues; and (iv) inhibitors of free fatty acid oxidation.


Subject(s)
Diabetes Mellitus, Type 2/therapy , Animals , Diet , Exercise , Humans , Hypoglycemic Agents/therapeutic use , Life Style , Weight Loss
16.
J Gerontol A Biol Sci Med Sci ; 52(5): B245-9, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9310073

ABSTRACT

Glucose intolerance is a common feature of the aging process, and aging per se is an etiologic factor for Type II diabetes mellitus. To characterize the beta cell abnormalities that occur with aging, we looked at the serum glucose and insulin levels of six young (3-month) and six old (22-month) Wistar rats at 0, 2, 4, 7, 10, 15, 20, and 30 minutes after an intravenous glucose load (IVGTT, 0.5 g/kg glucose). We found that the fasting glucose and insulin levels were not significantly different between young and old rats. However, peak glucose levels were significantly higher in the old (349 +/- 10 mg/dl) compared to the young (250 +/- 7 mg/dl) animals (p < .0001). Insulin levels in the young animals peaked at 2 minutes (859 +/- 171 pmol/l) with a quick return toward fasting levels by 7 minutes. The old animals had a delayed and blunted insulin response to glucose, achieving lower peak insulin levels (656 +/- 164 pmol/l) 7 minutes after the glucose load. As insulin levels are also positively modulated by incretin hormones, we quantitated the fasting insulin responses of young and old animals to .05, 0.1, 0.2, 0.4, and 0.5 nmol/kg intravenous glucagon-like peptide-1 (GLP-1), the most potent incretin known. Insulin responses were similar in both age groups, with maximum insulin responses seen at 0.4 nmol/kg. GLP-1, in conjunction with the IVGTT, restored the acute insulin response to glucose and increased the clearance of glucose in the old animals. It therefore appears that old animals have an impaired glucose-mediated insulin release but maintain their insulin responsivity to GLP-1. This makes it a likely candidate in the treatment of Type II diabetes.


Subject(s)
Aging/metabolism , Glucose/pharmacology , Insulin/metabolism , Peptides/pharmacology , Animals , Blood Glucose/analysis , Dose-Response Relationship, Drug , Fasting , Glucagon-Like Peptide 1 , Glucose Tolerance Test , Insulin Secretion , Male , Rats , Rats, Wistar
17.
J Clin Invest ; 99(12): 2883-9, 1997 Jun 15.
Article in English | MEDLINE | ID: mdl-9185511

ABSTRACT

Wistar rats develop glucose intolerance and have a diminished insulin response to glucose with age. The aim of this study was to investigate if these changes were reversible with glucagon-like peptide-1 (GLP-1), a peptide that we have previously shown could increase insulin mRNA and total insulin content in insulinoma cells. We infused 1.5 pmol/ kg-1.min-1 GLP-1 subcutaneously using ALZET microosmotic pumps into 22-mo-old Wistar rats for 48 h. Rat infused with either GLP-1 or saline were then subjected to an intraperitoneal glucose (1 g/kg body weight) tolerance test, 2 h after removing the pump. 15 min after the intraperitoneal glucose, GLP-1-treated animals had lower plasma glucose levels (9.04+/-0.92 mmol/liter, P < 0.01) than saline-treated animals (11.61+/-0.23 mmol/liter). At 30 min the plasma glucose was still lower in the GLP-1-treated animals (8.61+/-0.39 mmol/liter, P < 0.05) than saline-treated animals (10.36+/-0.43 mmol/liter). This decrease in glucose levels was reflected in the higher insulin levels attained in the GLP-1-treated animals (936+/-163 pmol/liter vs. 395+/-51 pmol/liter, GLP-1 vs. saline, respectively, P < 0.01), detected 15 min after glucose injection. GLP-1 treatment also increased pancreatic insulin, GLUT2, and glucokinase mRNA in the old rats. The effects of GLP-1 were abolished by simultaneous infusion of exendin [9-39], a specific antagonist of GLP-1. GLP-1 is therefore able to reverse some of the known defects that arise in the beta cell of the pancreas of Wistar rats, not only by increasing insulin secretion but also by inducing significant changes at the molecular level.


Subject(s)
Aging/physiology , Blood Glucose/metabolism , Glucose Tolerance Test , Peptides/pharmacology , Animals , Gene Expression/drug effects , Glucagon-Like Peptide 1 , Glucokinase/genetics , Glucose/administration & dosage , Glucose Transporter Type 2 , Insulin/blood , Insulin/genetics , Insulin/metabolism , Monosaccharide Transport Proteins/metabolism , Pancreas/drug effects , Pancreas/metabolism , Peptides/blood , RNA, Messenger/metabolism , Rats , Rats, Wistar
18.
Endocrinology ; 138(5): 1829-35, 1997 May.
Article in English | MEDLINE | ID: mdl-9112375

ABSTRACT

In this study, a partial hamster complementary DNA encoding ERCC-1, a member of the DNA excision repair gene family, has been cloned. The nucleic acid and amino acid sequences were highly homologous to those of human and mouse ERCC-1. The hamster ERCC-1 gene was expressed as a 1.2-kilobase message in cultured Chinese hamster ovary cells. Northern (RNA) blot analysis revealed that overexpression of the insulin receptor or various growth factor receptor tyrosine kinases in Chinese hamster ovary cells increased ERCC-1 messenger RNA (mRNA) levels. This effect did not occur in cells overexpressing mutated insulin receptors that are known to have impaired kinase-related signaling. Increased ERCC-1 expression correlated with resistance to UV exposure. Fluorescent-activated cell sorter analysis of confluent cell populations indicated no differences in cell cycle distribution. Furthermore, no significant relationship was demonstrated between the relative expression of ERCC-1 mRNA and the rate of glucose utilization. Insulin enhanced the accumulation of ERCC-1 mRNA in serum-deprived cells expressing wild-type insulin receptors. The potential role for activation of the insulin receptor and related growth factor receptors in ERCC-1 gene expression and function remains to be defined.


Subject(s)
DNA-Binding Proteins , Endonucleases , Gene Expression , Proteins/genetics , RNA, Messenger/metabolism , Receptor, Insulin/genetics , Receptor, Insulin/metabolism , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , CHO Cells , Cricetinae , DNA Repair , DNA, Complementary/chemistry , Flow Cytometry , Gene Expression/drug effects , Glucose/metabolism , Humans , Insulin/pharmacology , Mice , Molecular Sequence Data , Mutation , Proteins/chemistry , Ultraviolet Rays
19.
Ann Surg ; 225(3): 327-32, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9060590

ABSTRACT

BACKGROUND AND OBJECTIVE: Factors that control pancreatic regenerating (reg I) gene expression are unknown, but it is believed that its expression may correspond with cellular differentiation. The authors recently demonstrated that reg I is expressed in AR42J, a rat acinar cell line whose state of differentiation can be modulated by dexamethasone. They used this line to study reg I expression during cellular proliferation and differentiation. METHODS: After treatment of cells with 10 nmol/L dexamethasone, proliferation was assayed by thymidine incorporation; differentiation by expression of elastase I mRNA. Reg I mRNA levels were measured using a rat reg I cDNA probe, and reg I protein levels assayed by enzyme-linked immunosorbent assay of cellular lysates with a polyclonal antibody. The effect of gastrin, cholecystokinin and glucagon on reg I expression was also studied. RESULTS: When compared with controls, treatment with dexamethasone caused thymidine incorporation to decrease and elastase mRNA levels to increase. Reg I mRNA decreased from controls of 100 +/- 16% to 40 +/- 18% (p < 0.05), and reg I protein levels decreased as well. Gastrointestinal hormones had no significant effect on either elastase or reg I gene expression. CONCLUSIONS: Expression of reg I inversely correlates with the level of cellular differentiation, can be modulated via the glucocorticoid receptor, and is a potential marker of gastrointestinal epithelial differentiation. Despite its presence within a pancreatic acinar cell line, reg I gene expression is not modulated by gastrointestinal hormones.


Subject(s)
Gene Expression Regulation , Genes, Regulator/genetics , Pancreas/cytology , Animals , Cell Differentiation , Rats
20.
J Mol Endocrinol ; 19(3): 241-8, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9460645

ABSTRACT

Glucagon-like peptide-1 (7-36 amide) (GLP-1) is known to increase insulin release when given as a bolus in the fasted and fed state. GLP-1 also increases glucose uptake and lipid synthesis in cultured adipocytes. In this study we investigated the effects of GLP-1 on glucose uptake and on the levels of expression of the facilitative glucose transporters, GLUT1 and GLUT4, in fully differentiated 3T3-L1 adipocytes. Cells were incubated with GLP-1 (10 nM) with or without insulin (10 and 100 nM) for 24 h. Under these conditions, GLP-1 alone caused an increase in basal and acute insulin-stimulated glucose uptake along with an increase in GLUT1 and GLUT4 protein levels. However, there was no change in the expression of GLUT1 and GLUT4 mRNAs. In the absence of GLP-1, prolonged exposure to insulin caused a marked reduction in the levels of GLUT4 mRNA and protein, and an inhibition of glucose uptake after an acute exposure to insulin. This insulin-induced down-regulation of GLUT4 was prevented when GLP-1 was present during the 24-h treatment. In contrast, the acute insulin-stimulated glucose uptake could not be restored by GLP-1. GLP-1 is therefore the first gut hormone shown to be capable of modulating glucose transporter levels in cultured adipocytes.


Subject(s)
Adipocytes/cytology , Adipocytes/metabolism , Hexoses/pharmacokinetics , Monosaccharide Transport Proteins/genetics , Muscle Proteins , 3T3 Cells , Animals , Cyclic AMP/metabolism , Deoxyglucose/pharmacokinetics , Gene Expression Regulation , Glucagon/pharmacology , Glucagon-Like Peptide 1 , Glucose Transporter Type 1 , Glucose Transporter Type 4 , Hypoglycemic Agents/pharmacology , Insulin/metabolism , Insulin/pharmacology , Mice , Monosaccharide Transport Proteins/drug effects , Monosaccharide Transport Proteins/metabolism , Peptide Fragments/pharmacology , Protein Binding , Protein Precursors/pharmacology , RNA, Messenger/drug effects , RNA, Messenger/genetics , Receptor, Insulin/genetics , Receptor, Insulin/metabolism
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