The relationship between sudomotor function and skin microvascular reactivity in individuals with type 1 diabetes of long duration.

AIM: The aim of this study was to assess the relationship between sudomotor function and microvascular perfusion in patients with type 1 diabetes (DM1). METHODS: We evaluated 415 patients (206 women), with DM1, median age of 41 (IQR: 33-53) years, disease duration of 25 (IQR: 20-32) years. We assessed metabolic control of diabetes and the presence of peripheral and cardiac autonomic neuropathy. Sudomotor function was assessed using Sudoscan device by electrochemical skin conductance (ESC). Microvascular function was measured by laser-Doppler flowmetry with basal perfusion, the peak flow after occlusion (PORHpeak) and THmax which is the percentage change between basal perfusion and the peak flow during thermal hyperemia (TH). The accumulation of advanced glycation end products in the skin was assessed by skin autofluorescence (AF) measurement using AGE Reader. We subdivided patients based on the presence of diabetic peripheral neuropathy (DPN), cardiac autonomic neuropathy (CAN) and according to normal value of ESC. RESULTS: Patients with abnormal ESC had higher skin AF [2.5 (2.1-2.9) vs 2.1 (1.9-2.5) AU, p < 0.001], lower eGFR [83 (72-96) vs 98 (86-108) ml/min/1.73 m2, p < 0.001], higher basal perfusion [25 (12-81) vs 14 (7-43) PU, p < 0.001], lower THmax [664 (137-1461) vs 1115 (346-1933) %, p = 0.002], higher PORHpeak [104 (59-167) vs 70 (48-135) PU, p < 0.001] as compared to subjects with normal ESC results. We found negative correlation between THmax and TG level (Rs = -0.14, p < 0.005), AF (Rs = -0.19, p = 0.001), vibration perception threshold - VPT (Rs = -0.24, p < 0.001) and positive correlation with HDL level (Rs = 0.14, p = 0.005), Feet ESC (Rs = 0.21, p < 0.001) and Hands ESC (Rs = 0.14, p = 0.004). We found positive correlation between PORHpeak and TG level (Rs = 0.14, p = 0.003), skin AF (Rs = 0.29, p < 0.001), VPT (0.27, p < 0.001) and negative correlation with eGFR (Rs = -0.2, p < 0.001), HDL (Rs = -0.12, p = 0.01), Feet ESC (Rs = -0.27, p < 0.001) and Hand ESC (Rs = -0.16, p = 0.002). CONCLUSION: Impaired microvascular reactivity is associated with sudomotor dysfunction in patients with type 1 diabetes.

Smoking and other factors associated with short-term partial remission of Type 1 diabetes in adults.

AIMS: The duration of partial remission of Type 1 diabetes is associated with the degree of initial metabolic disturbance and features of insulin resistance. Cigarette smoking decreases insulin sensitivity, but its influence on the length of remission is unknown. Therefore, this study assessed the relationship between cigarette smoking and duration of partial remission in adults with newly diagnosed Type 1 diabetes. METHODS: We recruited 149 patients (48 women and 101 men, aged 16-35 years, median age 25 years), admitted to a teaching hospital with newly diagnosed Type 1 diabetes and followed them for a median period of 1 year and 9 months. We introduced intensive insulin therapy in multiple injections (basal-bolus) in all patients. We defined partial remission as an insulin dose of ≤ 0.3 U/kg body weight/24 h, an HbA(1c) value < 53 mmol/mol (7.0%) and a random serum C-peptide concentration over 0.5 ng/ml. Cigarette smoking was determined by self-report. RESULTS: Of 149 patients, 68 (46%) fulfilled the criteria for partial remission at 1 year after diagnosis of diabetes. Fewer patients who were in partial remission at 1 year smoked (19/68, 28%) than did patients that were not in partial remission (41/81, 51%). In logistic regression analyses, non-smoking was associated with remission at 1 year independent of age, sex, HbA(1c) and presence of diabetic ketoacidosis, all measured at onset of diabetes (OR 3.32, 95% CI 1.42-7.75, P = 0.005). CONCLUSION: Relative to individuals in this study who smoked, those who did not smoke at diagnosis of Type 1 diabetes experienced a longer duration of partial remission.

Evidence of polymorphonuclear neutrophils (PMN) activation in patients with insulin-dependent diabetes mellitus.

Polymorphonuclear neutrophils' chemotaxis, surface charge, superoxide anions generation, NBT (nitro blue tetrazolium) reduction and intracellular lysozyme, and beta-glucuronidase content were estimated in patients with type I diabetes mellitus in a similar state of metabolic control. The chemotaxis of diabetic cells toward bacterial chemotactic factors was similar to controls, whereas migration toward complement-derived chemoattractants was significantly reduced. Polymorphonuclear neutrophils isolated from diabetic patients, when unstimulated, produced significantly greater amounts of superoxide anions and reduced NBT more efficiently. They also revealed reduced surface charge and lower intracellular content of lysozyme, whereas beta-glucuronidase content was similar to controls. The results obtained seem to indicate that neutrophils in patients with insulin-dependent diabetes manifest signs of being in the activated state. The possible mechanisms of such stimulation are discussed.

Possible prognostic value of plasma activity augmenting polymorphonuclear neutrophils (PMN) adherence estimations in untreated patients with chronic myeloid leukemia.

Plasma activity augmenting polymorphonuclear neutrophils adherence was evaluated in 30 untreated patients with chronic myeloid leukemia. The estimations were done by the use of cells isolated from healthy persons. The plasma of some patients with CML induced the intensive adherence of PMN to plastic surface. Further observations suggested, that these patients were characterized by earlier blast crisis occurrence and poorer prognosis.

Polymorphonuclear neutrophils adherence, superoxide anion (O2-) production and HBA1 level in diabetic patients.

It was established that metabolic disturbances accompanying the impairment of diabetes control induced polymorphonuclear neutrophil (PMN) activation. Therefore, we evaluated the influence of diabetes control, measured as HbA1, level on superoxide anions production and neutrophils adherence to foreign surface. Twenty patients with insulin-dependent or non-insulin-dependent diabetes being in different state of metabolic control were studied. It was revealed that PMN from diabetics demonstrated significantly increased unstimulated superoxide anion production (11.07 +/- 1.25 vs. 4.15 +/- 1.24 nmol O2-/10(6) PMN/10 min). Stimulated O2- production did not reach the same levels as controls (21.80 +/- 1.34 vs. 23.67 +/- 4.36 nmol O2-/10(6) PMN/10 min). There were highly significant correlations between unstimulated O2- production and HbA1, levels (r = 0.86). Adherence of diabetic PMN suspended in autologous plasma was significantly increased in comparison to controls (12.76 +/- 1.20 vs. 6.04 +/- 1.99 cells/mm2). However, these data did not correlate with HbA1 level (r = 0.06). In conclusion our results seems to indicate that metabolic control of diabetes influence PMN function.

Impairment of the oxygen-dependent microbicidal mechanisms of polymorphonuclear neutrophils in patients with type 2 diabetes is not associated with increased susceptibility to infection.

Phagocytosis, bactericidal capacity and some selected parameters of oxygen-dependent bactericidal mechanisms were evaluated in 20 patients with type 2 diabetes being in similar (intermediate) state of metabolic control and in 15 healthy individuals. Polymorphonuclear neutrophils (PMNs) from diabetics showed normal ability to phagocytose staphylococci, a decreased Intracellular bacteria killing, the impaired stimulated superoxide anion (O2-) and hydrogen peroxide (H2O2) production and the low intracellular myeloperoxidase activity. The obtained data seem to indicate that the decreased bacterial killing by PMNs isolated from diabetics are partly at least related to an impairment of the oxygen-dependent bactericidal mechanisms. Since none of the diabetic patients suffered from recurrent infection the clinical significance of our finding is still uncertain.

Metabolic control quality and free radical activity in diabetic patients.

The role of oxidative/reductive balance derangement in the pathogenesis of diabetic microangiopathy has often been discussed in the last few years. Therefore, we decided to evaluate the influence of intensive insulin therapy on selected indicators of free radical production. The levels of plasma hydrogen peroxide (H2O2) and serum malonyldialdehyde (MDA) were estimated in 15 patients with Type 1 and 15 with Type 2 diabetes before and after 2 weeks of intensive treatment. The initial H2O2 and MDA levels in all cases were significantly higher than in controls. After 2 weeks of treatment, the values for both estimated parameters were significantly lower; however, they were still higher than in the control group. Our results seem to confirm the previous suggestions concerning the relation between metabolic disturbances and oxidative stress in diabetic patients.

The influence of insulin-dependent diabetes mellitus (IDDM) duration on superoxide anion and hydrogen peroxide production by polymorphonuclear neutrophils.

We address the question whether oxygen metabolism of polymorphonuclear neutrophils (PMN) is influenced by disease duration in patients with insulin-dependent diabetes mellitus (IDDM). PMN were isolated from patients with IDDM of various durations and from healthy controls. We measured PMN production of superoxide anions (O2-) by cytochrome c reduction (see Babior, B.M. et al. (1973) J. Clin. Invest. 52, 741-746) and PMN production of hydrogen peroxide (H2O2) by phenol red oxygenation (see Pick, E. (1980) J. Immunol. Methods 38, 161-169) in three groups of IDDM patients subdivided according to disease duration (group A: IDDM less that 10 years; group B: IDDM of 10-15 years; group C: IDDM of more than 15 years) and in control healthy subjects (group H). Unstimulated O2- production in all IDDM patients was not statistically different from control values (A: 4.3 +/- 0.4 nmol/10(6) PMN per 30 min, nmol/10(6) PMN per 30 min; C: 4.9 +/- 0.9 nmol/10(6) PMN per 30 min; and H: 3.5 +/- 0.2 nmol/10(6) PMN per 30 min, respectively). In contrast, stimulated O2- production was significantly lower in both patients with 10-15 years, and patients with more than 15 years, duration of IDDM than in controls (B: 25.7 +/- 2.5 nmol/10(6) PMN per 30 min; C: 21.1 +/- 3.4 nmol/10(6) PMN per 30 min and H: 42.2 +/- 1.1 nmol/10(6) PMN per 30 min, respectively) correlating with disease duration (r = -0.44, P < 0.033). The stimulated O2- production in patients with less than 10 years duration of IDDM (A: 35.7 +/- 1.9 nmol/10(6) PMN per 30 min) was slightly lower than in controls. H2O2 production of unstimulated PMN (A: 4.0 +/- 0.5 nmol/10(6) PMN per 30 min; B: 4.4 +/- 0.8 nmol/10(6) PMN per 30 min and C: 4.4 +/-1.0 nmol/10(6) PMN per 30 min, respectively) was much higher than those in controls. In contrast, stimulated H2O2 production did not differ statistically from the value noticed in healthy subjects. The results obtained might indicate that production of H2O2 by unstimulated cells is increased in diabetic patients while generation of O2- by stimulated neutrophils is markedly impaired, suggesting that toxic oxygen species production might be influenced by disease duration.

Hematological effects of the new immunosuppressive drug 15-deoxyspergualin.

Since systematic hematological studies on blood and bone marrow changes after treatment with 15-Deoxyspergualin (DOS) are lacking, a quantitative assessment was performed fourteen or twenty eight days after intraperitoneal application of DOS to rats. Further observations done 7 and 14 days after discontinuation of DOS administration allowed analysis of bone marrow regeneration. DOS induced lymphocytopenia, granulocytopenia and anemia with a decrease of bone marrow cellularity due to suppression of cell maturation. The effect was dose-dependent and bone marrow as well as blood changes were observed in animals treated with doses from 0.5 to 10.0 mg/kg DOS. Within 14 days after termination of the treatment, rapid recovery with normalization of all hematological parameters was observed. In the light of our data, these hematological side effects may not be a major disadvantage, if DOS is used in doses below 2.5 mg/kg, and for a course of therapy which is limited to 7 to 14 days.

Intravascular degranulation of polymorphonuclear neutrophils during hemodialysis. The effects of pretreatment with colchicine.

Polymorphonuclear neutrophil (PMN) counts, plasma activity inducing PMN aggregation, augmenting PMN adherence and chemotactic activity were estimated during hemodialysis (HD) with cuprophane membranes. All these plasma activities are accepted as related to the presence of activated complement (C). The same estimations were made for hemodialyzed patients pretreated with colchicine. The results, especially comparison of plasma activities in blood entering and leaving the dialyzer, indicate that significant amounts of activated C components are generated within the patients vascular bed. The PMN entrapped in the pulmonary microcirculation that undergo subsequent degranulation are most probably the source of these C components. This concept was confirmed by the results for patients pretreated with colchicine, who had significantly less of the activities generated in the patients own circulation.

Cardiopulmonary bypass-induced complement activation. Evidence for C5a participation.

Plasma activity inducing polymorphonuclear neutrophils (PMN) aggregation, augmenting PMN adherence, plasma chemotactic activity as well as peripheral PMN count were estimated in patients during cardiopulmonary bypass. In the course of the surgical procedure the chemotactic activity and the activity inducing PMN aggregation were detected in plasma. The top of these two activities was always followed by the significant fall of the peripheral PMN count. The increase of the activity augmenting PMN adherence was observed in a part of cases, but when noticed, always preceded the maximum of the remaining evaluated activities.

[Effect of immunomodulators on selected functions of neonatal polymorphonuclear neutrophils in vitro]. / Wplyw immunomodulatorów na wybrane funkcje granulocytów obojetnochlonnych noworodka in vitro.

Phagocytosis, bactericidal capacity, production of superoxide anions O2- and expression of receptor for the Fc portion of IgG of polymorphonuclear neutrophils (PMN) were estimated in umbilical venous blood of neonates and their mothers. We also evaluated the influence of immunomodulating agents on these parameters. The influence of the maternal serum on the function of neonatal PMN was examined. The obtained results indicate that functional defect of neonatal PMN may be corrected under the influence of the maternal serum and/or immunomodulating agents.

[Phagocytosis and bactericidal capacity of polymorphonuclear neutrophils in neonates]. / Fagocytoza i zdolnosc bakteriobójcza granulocytów obojetnochlonnych noworodków.

Phagocytosis and bactericidal capacity of polymorphonuclear neutrophils (PMN) obtained from umbilical venous blood was estimated in 30 neonates and their mothers by the use of fluorochrome microassay of Pantazis and Kniker. Phagocytosis of Staphylococcus was similar in both groups and controls, while intracellular bacteria killing was significantly impaired in PMN obtained from the neonates. These results may indicate that increased susceptibility to infection observed in neonates may be partly caused by PMN function impairment.

[Evaluation of selected components in antioxidant systems of blood in patients with diabetes]. / Ocena wybranych ukladów antyoksydacyjnych we krwi chorych na cukrzyce.

Oxidative stress plays a major role in the development of chronic complications of diabetes. The aim of our study was to evaluate the selected components of the antioxidative system in well metabolically controlled diabetic patients. We also decided to assess the correlation between these parameters and duration of disease and the presence of it's late complications. The study was entered by 30 patients with type 1 diabetes (18 female and 12 male, aged 30.2 + 10.8 years with mean duration of disease 8.37 + 6.56 years, HbA1c 6.8 + 1.6%). 24 healthy, sex- and age-matched volunteers served as controls. We assessed the following parameters: reduced glutathione in erythrocyte lysate (colorimetric method by Bioxytech GSH-400), serum glutathione peroxidase (enzymatic immunological method by Bioxytech pl. GPx-EIA) and plasma superoxide dismutase activity (colorimetric method based on cytochrome c reduction). In comparison with controls, we found significantly higher reduced glutathione level (11.20 + 0.79 vs 3.92 + 0.62 mumol/l, p < 0.001) and markedly lower dismutase activity (27.49 + 1.32 vs 39.73 + 4.45 U/ml, p < 0.001). The levels of glutathione peroxidase did not differ significantly from values obtained in healthy subjects. We did not observe any correlation between the analysed parameters and duration of diabetes, HbA1c or presence of chronic complications of disease. The obtained results might indicate that antioxidative systems in the state of good metabolic control of diabetes have adaptive properties.

[Evaluation of glucose, insulin, C-peptide uric acid serum levels in patients with psoriasis]. / Ocena stezenia glukozy, insuliny, peptydu C i kwasu moczowego w surowicy chorych na luszczyce pospolita.

The aim of our study was to estimate the serum levels of glucose, insulin, C-peptide and uric acid in patients with psoriasis before and after treatment. The study included 12 males with active form of psoriasis and 15 control subjects carefully matched to the psoriatic patients for age and BMI. All measured parameters were in patients with psoriasis significantly increased and dependent on the BMI. Compared with pretreatment values of glucose and uric acid were significantly lower during therapy. The increase in the mean C-peptide and insulin levels after psoriasis therapy was constant and independent from clinical stage of disease. The results of the present study have provided evidence for the importance of impaired glucose and purine metabolism in patients with psoriasis in the increase risk of development of diabetes mellitus and hypertension.

[Effect of prednisone and typhoid vaccine on the size of systemic neutrophil granulocyte pool]. / Wplyw prednisonu i szczepionki durowej na wielkosc ustrojowej puli granulocytów obojetnochlonnych

Changes of total granulocyte pool in the organism were evaluated after administration of prednisone or typhoid vaccine. For this purpose the unsaturated capacity of serum vitamin B12 binding was determined. After administration of prednisone the rise of granulocytes in peripheral blood was associated with a rise in the unsaturated capacity of vitamin B12 binding by the serum which was not observed after administration of typhoid vaccine. This observation confirmed the suggestions that corticosteroids not only mobilize the bone marrow reserve of granulocytes but prolong also their survival in the organism.

[Tissue migration of neutrophil granulocytes in the course of antiproliferative treatment in Hodgkin's disease]. / Migracja tkankowa granulocytów obojetnochlonnych w czasie leczenia antyproliferacyjnego ziarnicy zlosliwej

The ability of neutrophil granulocytes to mobilization in sterile inflammatory focus was studied in patients with Hodgkin's disease during treatment. Plastic chambers filled with patient's serum were used for this purpose. The migration ability of granulocytes in the group of patients was reduced already before beginning of treatment. It decreased even more 3 days after beginning of cytostatic treatment but on the 7th day the number of cells passing into the inflammatory exudate increased to values observed in healthy subjects. The observed changes were not correlated with fluctuations in the granulocyte count in the peripheral blood observed on the same days. It is suggested that the evaluation of migration ability of granulocytes to the inflammatory focus may provide better information about the defensive mechanisms of the organism than determination of the count of these cells in the peripheral blood.

[Lysozyme activity (LZM) and unsaturated vitamin B-12-binding capacity (NZW vit B-12) in the serum following prednisone administration in patients with bone marrow aplasia with pancotypenia]. / Zachowanie sie aktywnosci lizozymu (LZM) i nienasyconej zdolnosci wiazania witaminy B12 (NZW wit. B12) w surowicy pod wplywem prednisonu u chorych z aplazja szpiku z pancytopenia.

In 18 patients with bone marrow aplasia with pancytopenia lysozyme activity and unsaturated vitamin B12-binding capacity in the serum were determined. These investigations, together with determinations of peripheral blood polymorphonuclear count, were done again after prednisone administration. In all cases a significant fall was found in the NZW vit B12 and LZM activity in the serum. A slight rise in the polymorphonuclear count in the 24th hour of the study was associated with a rise in the NZW wit. B12 in the serum, and decreased LZM activity. This confirmed the previously demonstrated complex character of corticosteroid action on the system of polymorphonuclears. These results point also to the usefulness of determination of unsaturated vitamin B12-binding capacity for evaluating the value of the total granulocyte pool in granulocytopenia.

[Usefulness of hydrocortisone test for assessment of bone marrow reserve in neutrophils]. / Przydatnosc testu hydrokortzonowego dla oceny wielkosci rezerwy szpikowej granulocytów obojetnochlonnych.

The activity of lysozyme and unsaturated binding capacity of vitamin B12 in the serum were determined in hydrocortisone test for calculating the reserve of granulocytes in bone marrow. A significant rise in the count of neutrophils in peripheral blood 3 hours after the beginning of the test was not associated with statistically significant changes in the determined parameters. This in an evidence that prolongation of the survival of cells in the circulation as one of the mechanisms of corticosteriod action on the granulocyte system is without any greater importance. This is connected, most probably, with short duration of the test. The rise of neutrophil count in peripheral blood after hydrocortisone administration may be regarded, therefore, as a measure of the value of one marrow reserve of these cells.