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ABSTRACT: Glucose metabolism disorders influence anticarcinogenic function of natural killer (NK) cells. The aim of this study was to evaluate the number and cytotoxic activity of NK cells in type 2 diabetic (T2D) patients with negative family history of cancer, type 2 diabetic subjects with newly diagnosed untreated colon cancer (T2DCC) and patients without type 2 diabetes with newly diagnosed, untreated colon cancer (CC). Incubation tests were performed in 18 T2D patients, treated with diet and oral antidiabetic agents, 16 T2DCC; cT1-4N0M0 (c-clinical diagnosis based on computed tomography, colonoscopy and histopathology) treated with diet and oral antidiabetic agents and 16 normoglycemic CC; cT1-4N0M0. Control group included 18 metabolically healthy (with normal fasting glucose and normal glucose tolerance) subjects (HS) with negative family history of cancer, matched by age, BMI and waist circumference. Peripheral blood mononuclear cells were isolated by means of gradient centrifugation. The K562 human erythroleukemia cell line served as the standard target for human NK cytotoxicity assay. The T2D revealed an increased number of NK cells (13.56 ± 5.9 vs 9.50 ± 4.8 %; p < 0.05) when compared with HS, yet these cells had a decreased activity (3.3 ± 2.5 vs 9.4 ± 3.6 %; p < 0.01). The CC demonstrated a decreased activity (2.9 ± 1.8 %; p < 0.01) but a similar number (8.82 ± 3.7 %; not significant) of NK cells when compared to HS. The T2DCC NK cells were characterized by trace cytotoxic activity (1.1 ± 0.7 %; p < 0.01) and nearly three times greater amount (21.24 ± 7.5 %; p < 0.01) when compared to T2D. Type 2 diabetes and CC are associated with disadvantageous alterations of NK cells, leading to impairment in their cytotoxic activity. The impaired activity of NK cells in T2D can be involved in the increased carcinogenic risk and can promote a higher incidence of CC.
Archivum Immunologiae et Therapiae Experimentalis 03/2013; · 2.54 Impact Factor
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ABSTRACT: The objective of this study was to evaluate glucose transport into lymphocytes in healthy subjects and patients with type 2 diabetes mellitus (DM) treated either with diet only or with insulin and to propose peripheral blood lymphocytes as a convenient model for cellular glucose transport studies.
Sixty subjects with type 2 DM, 30 treated with diet only and 30 with insulin, were investigated. Thirty healthy subjects matched for age, weight, and sex served as a control group. Deoxy-D-glucose, 2-[(3)H(G)] transport was studied in isolated peripheral blood lymphocytes. Expression of glucose transporters was ascertained by immunocytochemical identification and by Western blotting.
In lymphocytes from the control group, deoxy-D-glucose uptake increased gradually with the duration of the experiment. In diabetics treated with insulin, the maximal increase in deoxy-D-glucose uptake was observed after 30 min of the investigation, followed by a plateau phase. In diabetics treated with diet, deoxy-D-glucose uptake increased slowly during the first 30 min. The presence of GLUT1 and GLUT3 in lymphocytes was confirmed in this study.
Glucose transport into lymphocytes is altered in type 2 DM. In lymphocytes from diabetics, the dynamics of deoxy-D-glucose uptake significantly differed from that in healthy subjects. There was also a significant difference between the diabetic groups, representing different modes of therapy and stages of the disease. Glucose transport into lymphocytes is apparently influenced by DM as well as by the mode of therapy. We suggest that peripheral blood lymphocytes may become a promising model for studies on glucose transport in diabetes.
Archivum Immunologiae et Therapiae Experimentalis 04/2012; 55(2):119-26. · 2.54 Impact Factor
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ABSTRACT: The proper expression of particular glucotransporter (GLUT) isoforms determines a sufficient supply of glucose to tissues. The impairment of cellular glucose transport observed in insulin resistance leads to glucose metabolism disturbances. The aim of this study was the estimation of insulin resistance indicators and the quantitative expression of GLUT-1, GLUT-3 and GLUT-4 on peripheral blood lymphocytes in prediabetic subjects and persons with a positive family history of type 2 diabetes during 24 months of observation.
The study included 25 prediabetic subjects (according to WHO criteria) and 24 normoglycaemic individuals with a positive family history of type 2 diabetes. Twenty three healthy subjects with no family history of type 2 diabetes, matched with BMI, served as a control group. All participants were recommended to perform physical activity for at least 140 minutes per week and to maintain a low calorie diet. The peripheral blood lymphocytes demonstrating expression of GLUT-1, GLUT-3 and GLUT-4 were labelled with the use of indirect immunofluorescence. The expression of GLUT isoforms was investigated by flow cytometry. Cells were stained by using anti-human GLUT antibodies and FITC-conjugated immunoglobulin. Flow cytometry was performed using a FACS Calibur (Becton-Dickinson). Additionally, we determined: fasting plasma glucose (FPG), insulin and C peptide concentrations, HOMA-IR, BMI and WHR. All the tests were performed at baseline, and after 12 and 24 months.
At baseline, prediabetics and subjects with a positive family history of type 2 diabetes were characterised by a much higher expression of GLUT-4 compared to control subjects. Twenty four months of lifestyle modification resulted in significant lowering of the expression of GLUT-4 on the surface of PBL in both studied groups, with no differences in the expression of GLUT-1 or GLUT-3. Both prediabetic subjects and individuals with a positive family history of type 2 diabetes revealed no significant differences in determined insulin resistance markers after 24 months of the observation compared to the baseline values.
The estimation of typical GLUT isoforms present on the peripheral blood lymphocytes, as well as the evaluation of insulin resistance indicators, are obviously insufficient for monitoring the metabolic disorders progression in the risk groups of type 2 diabetes. The decrease in GLUT-4 lymphocyte expression may reflect a positive influence of lifestyle modification on a tissue redistribution of this crucial insulin-dependent glucotransporter. The determination of GLUT-4 on the surface of peripheral blood lymphocytes can be a useful tool for the evaluation of the efficacy of therapeutic actions in subjects at high risk of type 2 diabetes.
Endokrynologia Polska 01/2012; 63(3):212-9. · 1.24 Impact Factor
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ABSTRACT: Diabetes and cancer are diseases which take the size of an epidemic spread across the globe. Those diseases are influenced by many factors, both genetic and environmental. Precise knowledge of the complex relationships and interactions between these two conditions is of great importance for their prevention and treatment. Many epidemiological studies have shown that certain types of cancer, especially gastrointestinal cancers (pancreas, liver, colon) and also the urinary and reproductive system cancers in women are more common in patients with diabetes or related metabolic disorders. There are also studies showing the inverse relationship between diabetes and cancer, or the lack of it, but they are less numerous and relate mainly to prostate cancer or squamous cell carcinoma of the esophagus. Epidemiological studies, however, do not say anything about the mechanisms of these dependencies. For this purpose, molecular research is needed on the metabolism of cells (including tumor cells) and on metabolic dysfunctions that arise due to changes in the cell environment taking place in the sick, as well as in the intensely treated human organism.
Archivum Immunologiae et Therapiae Experimentalis 03/2011; 59(3):215-30. · 2.54 Impact Factor
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ABSTRACT: The normalization of cellular glucose assimilation is the basic aim of metabolic therapy in type 2 diabetes mellitus (T2DM). It requires parallel changes in the process of cellular glucose transport (CGT). Therefore the level of CGT could be regarded as a therapeutic target for oral hypoglycemic drugs in T2DM. To explore this hypothesis, CGT levels before and after sulfonylurea therapy were investigated. Peripheral blood lymphocytes were used as a cell model for testing CGT.
CGT was assessed by experimental in vitro tests allowing timed comparative observation of the transport process during the incubation of lymphocytes with 2-[(3)H(G)] glucose under basal conditions and after the addition of sulfonylurea or sulfonylurea plus insulin. The incubation tests were performed at baseline in 28 persons with newly diagnosed, therapy-naive T2DM and in 20 control subjects. In the diabetic patients the tests for CGT were repeated after 3 months of sulfonylurea therapy. The level of glucotransporter 4 (GLUT4) expression was also assessed by flow cytometry before and after the therapy.
Before treatment, CGT was significantly lower in the subjects with T2DM. The cells responded to the addition of sulfonylurea by a moderate increase in CGT. This response was augmented by the addition of insulin to sulfonylurea in the culture medium.
The three-month therapy with sulfonylurea resulted in a significant increase in CGT in all types of culture tests. This sulfonylurea-related improvement in CGT was associated with a near normalization of GLUT4 expression in the cells.
Archivum Immunologiae et Therapiae Experimentalis 11/2009; 57(6):467-73. · 2.54 Impact Factor
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ABSTRACT: The aim of the study was in vitro evaluation of piroxicam solid dispersions containing hydroxypropyl methylcellulose acetate succinate (HPMCAS-LF, -HF) as a carrier. Binary (piroxicam-HPMCAS) and ternary (piroxicam-HPMCAS-Carbopol 940) solid dispersions were prepared by spray-drying method. The morphological characteristics were investigated by scanning electron microscopy. X-ray diffraction and differential scanning calorimetry were employed to study physical and chemical properties. In vitro release was studied using a flow-through cell technique. Studies of dissolution rate of piroxicam from solid dispersions were carried out in comparison with corresponding physical mixtures and drug alone. The dissolution profiles depend on the presence of Carbopol 940 in solid dispersions.
Pharmaceutical Development and Technology 09/2008; 13(6):495-504. · 1.36 Impact Factor
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ABSTRACT: Ischemic heart disease (IHD) and acute myocardial infarction (AMI) present several important specificities in their clinical course. The problem how these circumstances influence the life of the persons with diabetes (DM) in longer perspective is not well elaborated. Therefore the assessment of the medical history of persons with DM after first MI was undertaken based on analysis of the following endpoints: the recurrence of MI, hospitalization due to acute heart ischemic incidents, the procedure of coronary revascularization, appearance of stroke, death due to cardiovascular causes. INVESTIGATED GROUP AND METHODS: During the years 1996-2000 69 subjects with AMI were hospitalized from the area of the local, urban hospital. The medical documentation supplemented by the patient and family home reviews and data from death registry office for the period of 2 years since the hospitalization due to first AMI was analyzed. RESULTS: In the group under study the males aged between 61 and 70 years were preponderant - 20,3%, the largest females subgroup was in the age between 71 and 80 years - 17,4%. The first AMI was the revelatory of the previously not known DM in 30,4% of cases, 21,7% subjects had the diagnosis of DM made in the last 5 years before first AMI. The symptoms of IHD were present before first AMI in 49% of subjects. 82% of them had the Q type AMI in 23% localized in inferior, in 17% in anterior and in 16% in lower-posterior wall of the heart. The frequencies of other then hyperglycaemia risk factors were: arterial hypertension - 67%, obesity and overweight respectively 38% and 26%, smoking - 32%, dyslipidemia - 28%. The risk factors were often multiple in the form of metabolic syndrome. During the 2 years of the follow-up after first AMI, 34,6% of subjects died. In 72,2% of this subgroup the mortality was due to the cardiovascular syndromes - recurrent MI, cardiac failure, stroke and "generalized atherosclerosis". The rate of hospitalization caused by acute ischemic incidents was 25%, recurrent MI - 11,5%, cardiosurgical intervention - 5,8%, stroke - 3,8%. CONCLUSIONS: The mortality level of 34,6% during 2 years after first AMI due to new cardiovascular events is the measure of the specific unfavourable prognosis for diabetic persons after first AMI. It underlines the need of the implementation in this subgroup of the intensified cardio-diabetological secondary prevention plan.
Polskie archiwum medycyny wewnȩtrznej 02/2006; 115(1):29-36. · 1.37 Impact Factor
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ABSTRACT: The normalization of cellular glucose assimilation is the basic aim of metabolic therapy in type 2 diabetes mellitus (T2DM). It requires parallel changes in the process of cellular glucose transport (CGT). This review presents the pathophysiological and clinical outlines of CGT. Sequentially, the advances in the mechanisms and classification of CGT and their physiological and molecular base are described. The role of CGT pathogenetic significance in diabetes mellitus is stressed. Finally, the opinion is expressed that the CGT study is a potentially important approach to clinical interpretation of glucose metabolism disturbances and their pharmacotherapy.
Endokrynologia Polska 61(3):303-10. · 1.24 Impact Factor
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ABSTRACT: GLUT4 is a representative of the family of integral membrane proteins which facilitate glucose transport across cellular membranes. In the available literature there is no publication referring to the investigations of glucotransporter expression in pre-diabetic subjects. However, GLUT4 protein overexpression was shown in leukocytes of diabetic patients. The aim of this study was to compare GLUT4 quantitative expression in peripheral blood lymphocytes in type 2 diabetes mellitus risk groups to healthy subjects.
The study groups included 15 pre-diabetic subjects and 15 persons with normal glucose tolerance and positive family history of type 2 diabetes mellitus (first-degree relatives). As a control group, 15 healthy persons with no family history of diabetes were enrolled. The expression of GLUT4 on the surface of peripheral blood lymphocytes was investigated with the use of indirect immunofluorescence. Quantitative determination of GLUT4 was performed with the use of flow cytometry.
In the control group, GLUT4 expression amounted to 12 + or - 1.5% and was significantly lower in relation to both pre-diabetic subjects (18.2 + or - 8.8%; p = 0.008) and the positive family history group (17.9 + or - 9%; p = 0.001).
GLUT4 overexpression in subjects with positive family history of type 2 diabetes mellitus suggests that cellular glucose transport disturbances occur prior to hyperglycaemia. Determination of GLUT4 expression appears to be a possibly useful method of early detection in individuals at high risk of diabetes.
Endokrynologia Polska 61(3):269-74. · 1.24 Impact Factor
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ABSTRACT: The current world epidemic of type 2 diabetes mellitus results from two general groups of causative factors. One is the influence of strong pathogenetic environmental pressures - also described as negative civilizational influence - on the very large subpopulation, assessed at 30% of the total world population, which is genetically predisposed to react to this external stress with the symptoms of type 2 diabetes mellitus. Such a pathogenetic reaction is based on the appearance of cellular and organ resistance to insulin. A second factor involves the beta cells of the pancreatic islets and their dysfunction. For these reasons, studies on the aetiology of insulin resistance have significance, both theoretical and practical. There are many biological deviations that can produce cellular insulin resistance and underutilization of glucose. The mechanism that is always present is the decrease of cellular glucose transport. For this reason, it should be approached as a potential target for preventive and therapeutic actions. These pathophysiological and clinical circumstances were the motivation for presenting a review of cellular glucose transport pathophysiology, which contributes to the aetiology of insulin resistance, cellular underutilization of glucose, and type 2 diabetes mellitus. They underline the significance of cellular glucose transport as a target for prevention and therapy of type 2 diabetes mellitus and other insulin resistant conditions. This review presents comments about the influence on cellular glucose transport of diet, physical exercise, and pharmacotherapeutic agents, based on the authors' studies. The review could contribute to an innovative approach to the pathogenesis, prevention, and therapy of type 2 diabetes mellitus and other conditions related to insulin resistance.
Endokrynologia Polska 61(3):292-302. · 1.24 Impact Factor
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ABSTRACT: This review presents the advances in the molecular biology and the pathophysiology of insulin resistance with emphasis on disturbances in cellular glucose transport. New scientific information about the structure and function of glucotransporters from the GLUT4 and SLGT families underline their significance in endocrinopathies and metabolic disease pathogenesis as related to insulin resistance. The new discoveries in this area also contribute to a better understanding of the regulation of insulin receptor and post-receptor reactivity by hormones and by drugs. They refer to the regulation of glycaemia and to its disturbances in diabetes mellitus, particularly of type 2, to metabolic syndrome, and, in general, to the pathogenesis of many syndromes and clinical disturbances caused by insulin resistance. Impairment of cellular glucose transport may be one of the primary aetiological factors in this respect. Therefore, studies of cellular glucotransporters expression and function promise new clinical and pharmacotherapeutic developments. Progress in this area has already been transformed into many practical proposals which are improving clinical practice.
Endokrynologia Polska 61(4):388-94. · 1.24 Impact Factor
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ABSTRACT: We investigated the effects of insulin on glucose transport in human peripheral lymphocytes using flow cytometry. We hypothesized that lymphocytes could be used as tools to study insulin action at the cellular level and facilitate the investigation of mechanisms that lead to insulin resistance.
Blood was withdrawn from 25 healthy subjects. The expression of glucose transporter (GLUT) isoforms in plasma membrane and the rates of glucose transport were determined with and without insulin (10 to 100 mU/L). Anti-CD3 phycoerythrin monoclonal antibody was used for lymphocyte gating. GLUT1, GLUT3, and GLUT4 isoforms were determined after staining cells with specific monoclonal antibodies to GLUT1, 3, and 4. Glucose transport was monitored with deoxy-D-glucose, 2-[3H(G)] - 185-370 GBq.
Insulin increased the uptake of deoxy-D-glucose and the expression of GLUT1, GLUT3, and GLUT4 isoforms in the plasma membrane. The optimal effects were always reached at 50 mU/L of insulin with the increase in GLUT1, 3, and 4 expression of 12%, 44%, and 38%, respectively. Mean baseline values of deoxy-D-glucose uptake were 3409 ccpm at 15 min., 6587 ccpm at 30 min., and 12525 ccpm at 60 min. of investigation. The maximal uptake in insulin-stimulated conditions was reached with 50 mU/L of insulin and went up to 12450 ccpm at 15 min., 37482 ccpm at 30 min., and 37916 ccpm at 60 min. of investigation (p < 0.01).
Peripheral blood lymphocytes may become an interesting model system to study the effects of insulin on cellular glucose transport. Flow cytometry is suitable for this investigation and may be used as a method to estimate the influence of insulin on GLUTprotein translocation and the dynamics of glucose uptake by lymphocytes.
Endokrynologia Polska 61(2):182-7. · 1.24 Impact Factor
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ABSTRACT: The normalization of cellular glucose assimilation is the basic aim of therapy in diabetes mellitus. This process should be accompanied by a proportional increase of the cellular glucose transport (CGT). The level of CGT should react to therapy typically recommended in Type 2 diabetes mellitus (T2DM), composed of diet and sulfonylurea. In order to explore this clinically significant hypothesis, a clinical-experimental study was undertaken. Its aim was to determine the clinical pharmacotherapeutic significance of CGT measurements.
CGT testing was performed on peripheral blood lymphocytes. CGT was assessed with 2-[(3)H(G)] glucose: before, and after the addition of sulfonylurea or sulfonylurea plus insulin to the incubation medium. Tests were performed at baseline in 28 persons with newly diagnosed, "therapeutically naive" T2DM and in 20 control subjects. In diabetic patients the tests for CGT were repeated after 3 months of routine diet and sulfonylurea therapy. In addition, the level of GLUT4 expression was assessed by flow cytometry before and after this therapy.
Before treatment, CGT was significantly decreased in all subjects with T2DM. Incubated in-vitro cells responded directly to the addition of sulfonylurea with a moderate increase of CGT. This response was augmented by the addition of insulin to sulfonylurea in the incubation medium. The monitored three-month routine, controlled therapy with diet and sulfonylurea resulted in a significant increase of CGT process in all types of incubation tests.
The basal and reactive CGT is significantly decreased in lymphocytes of persons with T2DM before the introduction of therapy. Effective therapy with diet and sulfonylurea normalizes both types of CGT - basal and reactive. It is related to the near normalization of GLUT4 expression in the studied cells. This phenomenon may be used as a new marker for diabetes mellitus pharmacotherapy. (Pol J Endocrinol 2010; 61 (1): 75-81).
Endokrynologia Polska 61(1):75-81. · 1.24 Impact Factor
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ABSTRACT: The levels of glycosylated hemoglobin (Hb AI), intermittent glycemia and glycosuria over 24 hr, Mw index, fasting serum cholesterol and triglycerides, and 24-hr proteinuria were determined in 20 healthy subjects and 88 diabetics representing different clinical types of diabetes mellitus. In each of the subjects all the tests were carried out on a single day. The other investigations included endogenous creatinine clearance, ECG and ophthalmoscopic examination of the eye-fundus.The mean Hb AI levels in the “A” control group (up to 40 yr of age) and in the “B” control group (41–60 yr) were of the total hemoglobin concentration, respectively. A significant increase in Hb AI concentration was found in all the diabetic patients. The increase, independent of the subject's age, clinical type of diabetes and the therapy employed, was related to the degree of hyperglycemia.In Type I diabetes there was no positive correlation between Hb AI concentration on the one hand and fasting glycemia, the 24-hr profile of glycemia, glycosuria and Mw index on the other. The latter indices of diabetes mellitus control seem thus to differ in value and significance. In Type II diabetes, both newly-diagnosed and of long duration, treated with the sulfonylurea derivatives, a marked correlation was found between Hb AI level and fasting glycemia, the mean value of 8 glycemia determinations over 24-hr, Mw index and 24-hr glycosuria. In Type II diabetes treated with insulin a correlation was established between Hb AI and other findings, except fasting glycemia. Hence, in diabetes of a more stable course, as opposed to Type I diabetes, determination of Hb AI is of less value.Triglyceride concentration was correlated with Hb AI concentration only in Type II diabetes of long duration, treated with the sulfonylurea derivatives. A significant correlation between Hb AI and cholesterol concentrations was found only in newly-diagnosed Type I diabetes.A comparative analysis of the results obtained seems to suggest that Hb AI concentration is also an indicator of the effects of diabetes therapy over longer intervals.
Journal of Chronic Diseases.
