Janusz Myśliwiec

CD11a Expression and soluble ICAM-1 levels in peripheral blood in high-risk and overt type 1 diabetes subjects

A pair of correspondent adhesion molecules: LFA-1 (CD11aCD18) and ICAM-1 (CD54) was shown to be involved in autoimmune insulitis in animal models. Anti-LFA-1 or anti-ICAM-1 monoclonal antibodies administered in vivo had a very strong preventive effect on the development of spontaneous diabetes with a marked reduction of insulitis. On the other hand elevated levels of the soluble form of ICAM-1 (sICAM-1) were documented in subjects at risk for type 1 diabetes. Recently sICAM-1 was shown to play an immunoregulatory role as an inhibitor of islet insulitis. The aim of the present study was to evaluate CD11a + mononuclear cells (lymphocytes and monocytes) and soluble sICAM-1 levels in the peripheral blood of subjects with preclinical and overt type 1 diabetes to assess their role in the development of the autoimmune process and their possible associations with the humoral autoimmune markers. The study was carried out in three groups of subjects: 26 first degree relatives of type 1 diabetes patients (prediabetics) with the combinations of autoantibodies against pancreatic B cells (ICA, GADA, IA-2A, IAA), 22 patients with a recent onset of type 1 diabetes and age and sex-matched 24 healthy volunteers (control group). We observed an increased fluorescence intensity of CD11a on mononuclear cells in overt diabetes subjects and a positive correlation between CD11a fluorescence intensity on monocytes and ICA titre. The highest sICAM-1 levels we obtained in the peripheral blood in the prediabetics in comparison to patients with clinical diabetes and the healthy controls. We found a positive correlation between slCAM-1 and values of ICA, GADA and a total number of antibodies present. In conclusion our study suggests that LFA-1 and sCAM-1 play an important role in the pathogenesis of type 1 diabetes. The assessment of the CD11a bearing monocytes and sICAM-1 levels are potential markers of the preclinical stage of the autoimmune diabetes, but further prospective studies in high risk diabetes type 1 subjects are needed.

Nicotinamide inhibits enhanced in vitro production of interleukin-12 and tumour necrosis factor-α in peripheral whole blood of people at high risk of developing Type 1 diabetes and people with newly diagnosed Type 1 diabetes

Macrophages and T lymphocytes are the first cells to appear in pancreatic islets in the development of autoimmune diabetes. It has been suggested that cytokines released by monocytes/macrophages, including interleukin-1beta (IL-1beta), interleukin-12 (IL-12) and tumour necrosis factor-alpha (TNF-alpha) could have an initial role in islet B-cell damage. The aim of the present study was to estimate the effect of human insulin and nicotinamide on the levels of monocyte/ macrophage derived cytokines in the peripheral blood of humans at risk of Type 1 diabetes, and in patients with newly diagnosed Type 1 diabetes compared to healthy control subjects. The study was carried out on three groups of subjects: 20 first degree relatives of people with Type 1 diabetes (with two or more antibodies against pancreatic B-cell antigens); 22 patients with recent onset of Type 1 diabetes (duration of the disease 3-6 months); and 25 age- and sex-matched healthy subjects. Cytokine levels (IL-1beta, IL-12, and TNF-alpha) in the supernatants of whole blood cultures incubated with PHA alone (10 microg/ml), or PHA + human insulin (50 microg/ml), or PHA + nicotinamide (100 micromol/l) were quantified by ELISA. In the cultures with nicotinamide the concentration of IL-12 and TNF-alpha was significantly lower in the prediabetic group, diabetic patients, and the healthy controls than in the cultures with PHA only or with PHA + insulin. There were no significant differences in IL-1beta production in the cultures after incubation with the different stimuli in the studied groups and healthy controls. No significant influence of human insulin on macrophage/monocyte cytokines secretion in in vitro cultures of the peripheral blood was found. This suggests that nicotinamide could influence monocyte/macrophage function in peripheral blood by inhibiting production of IL-12 and TNF-alpha.

The Changes in the Endothelial Function and Haemostatic and Inflammatory Parameters in Subclinical and Overt Hyperthyroidism

Introduction. The aim of the present study was to compare the levels of circulating markers of endothelial function and low-grade inflammation in patients with subclinical and overt hyperthyroidism (OH) due to Graves disease (GD) and toxic nodular goiter (TNG). Material and Methods. The group studied consisted of 42 patients with GD, 75 patients with TNG, and 39 healthy controls. Results. Circulating markers of endothelial dysfunction were elevated in the patients with both SH and OH, but the concentrations of interleukin-12 (IL-12) (P < 0.05), IL-18 (P < 0.05), fibrinogen (P < 0.01), and von Willebrand factor (vWF) (P < 0.05) were significantly higher in the OH than in the SH group. The highest levels of IL-6, IL-12, IL-18, vWF, sVCAM-1, and fibrinogen were found in the patients with GD, but the differences between the GD, and TNG groups were not significant. In the subjects with OH serum IL-6 was positively associated with FT3 (R = 0.276, P < 0.05), FT4 (R = 0.273, P < 0.05), and thyroid peroxidase antibodies (R = 0.346, P < 0.01) levels. Conclusion. Our results may suggest that both SH and OH may be associated with endothelial dysfunction, which is reflected by decreased fibrinolytic activity, hypercoagulability, and increased levels of IL-6, IL-12, and IL-18 and depends not only on the cause but also on the degree of hyperthyroidism.

Diagnostics of primary aldosteronism: is obligatory use of confirmatory tests justified?

Introduction: Assessment of the renin-angiotensin-aldosterone system has been recently granted a much greater role in the evaluation of patients with arterial hypertension. There is no single test efficient in selection of patients for second-step etiological investigation. Methods: Altogether, 198 consecutive patients − 119 women (60%) and 79 men (40%) – hospitalized in years 2009–2011 at the Clinical Department of Endocrinology Medical University of Bialystok were diagnosed with primary aldosteronism. In each patient, plasma renin activity and plasma aldosterone concentration (basic and after 2 l NaCl infusion) were evaluated. Results: The percentage of patients with plasma aldosterone concentration ≥15 ng/ml was 53 and the percentage of patients with plasma renin activity ≤0.1 ng/ml/h was 20. The percentage of patients screened for primary aldosteronism in which the aldosterone:renin ratio exceeded consecutive cut-offs of 20, 30, 40 and 50 were respectively 57, 45, 34 and 29. Among 15 patients in which plasma aldosterone concentration after infusion of 2 l of saline was ≥6.5 ng/dl (8.6%), 13 (6.6%) were diagnosed with primary aldosteronism. Conclusion: The obligatory use of tests confirming autonomy of aldosterone secretion in patients screened for primary aldosteronism seems cost-effective in limiting the number of patients for further diagnosis.

Single, very low dose (0.03 mg) of recombinant human thyrotropin (rhTSH) effectively increases radioiodine uptake in the I-131 treatment of large nontoxic multinodular goiter

BACKGROUND Radioiodine therapy (RIT) in patients with large nontoxic multinodular goiter (MNG) recently becomes more common method in comparison to surgery (especially in elderly patients and with contraindications because of severe chronic diseases other systems). Repeatedly low thyroid radioactive iodine uptake (RAIU) decreases effectiveness of RIT or makes it impossible. The recombinant human thyrotropin can increase RAIU and improve the results of RIT. THE AIM OF THE STUDY was to assess the influence of a single very low dose (0.03 mg) of rhTSH on RAIU and thyroid function in euthyroid (MNG-EU) and subclinical hyperthyroid (MNG-SC) patients with a large multinodular goiter. MATERIAL AND METHODS 40 patients (14 male, 26 female, age 57-80 yr) with large non-toxic MNG over 80 grams and with baseline RAIU < 40% were included into the double-blind randomized study and divided into two groups: rhTSH-group and control group. First group received the single intramuscular injection of 0.03 mg rhTSH and the second received placebo. The RAIU were measured 24 and 48 hours after the rhTSH and then all the patients were administered therapeutic doses of I-131. TSH and free thyroxine levels were measured at 1st and 2nd day after the injection of rhTSH and later, at 4 and 8 weeks after the RIT. RESULTS The mean RAIU increased significantly from 30.44 ± 7.4% to 77.22 ± 8.7% (p < 0.001). There were no statistically significant differences in RAIU between euthyroid (MNG-EU) and subclinically hyperthyroid (MNG-SC) patients. The peak of serum TSH was noticed 24 hours after rhTSH injection and in MNG-EU patients it has remained within normal range, similarly as fT4. In the MNG-SC group the administration of rhTSH resulted in a significant increase in the TSH values after 24 hours, whose mean level slightly exceeded the upper limit of the normal range with normalization at 48 hours. 8 weeks after the RIT, the TSH and fT4 levels did not exceed the normal range and did not differ in a statistically significant way. CONCLUSIONS Even the single very low dose of rhTSH increases the values of RAIU in significant way, in euthyroid and subclinically hyperthyroid patients. The administration of rhTSH is well-tolerated. Neoadjuvant administration of a low dose (0.03 mg) of rhTSH before I-131 seems to be an optimal method of management which may increase the effectiveness of RIT and decrease the exposure of the patients to absorbed doses of ionizing radiation.

Calculation of therapeutic activity of radioiodine in Graves’ disease by means of Marinelli’s formula, using technetium (99mTc) scintigraphy

The therapeutic activity of 131I administered to patients with Graves’ disease can be calculated by means of Marinelli’s formula. The thyroidal iodine uptake (131IUmax) needed for the calculation is usually determined with the use of 131I. The purpose of the paper was to estimate 131IUmax on the basis of technetium uptake in the thyroid at 20 min (99mTcU20min). Eighty patients suffering from Graves’ disease were qualified for radioiodine therapy with measurement of fT4, fT3, thyroid-stimulating hormone and its receptor (TRAb). Prior to the treatment, all the patients were euthyroid. 131IUmax for each patient was determined according to the levels of 131I after 24 h (131IU24h), while effective half-life (Teff) according to the measurements of 131IU24h and 131I uptake after 48 h (131IU48h). Additionally, on the day before measuring 131IU24h, 99mTcU20min was calculated for each patient. It was demonstrated that there existed a correlation, with statistical significance at p < 0.05, between the following pairs of values: TRAb and 131IU24h, TRAb and 99mTcU20min, and 99mTcU20min and 131IU24h. The interdependence between 131IU24h and 99mTcU20min at the level of significance p < 0.05 is described by the following algorithms: 131IU24h = 17.72 × ln (99mTcU20min) + 30.485, if TRAb < 10 IU/ml, and 131IU24h = 18.03 × ln (99mTcU20min) + 38.726, if TRAb > 10 IU/ml. It is possible to predict thyroid iodine uptake 131IU24h in Graves’ disease on the basis of measuring the uptake of 99mTcU20min. This shortens the time necessary for diagnosis and enables the calculation of 131I activity using Marinelli’s formula.

Diagnostics of hypercortisolism — comparison between the clinical usefulness of salivary and serum cortisol measurements

INTRODUCTION The aim of this study was the comparison of 24h urine free cortisol (UFC), serum cortisol at 11pm (SCM) and late-nightsalivary cortisol (LSC) in patients suspected for hypercortisolism, and an assessment of the usefulness of these measurements in diagnosingovert Cushings (OCS) syndrome, pseudo Cushings state (PCS) and subclinical Cushings syndrome (SCS). MATERIAL AND METHODS The study group consisted of 82 patients, of whom four patients had SCS, three OCS and eight PCS. For measurementsof LSC, the ELISA method was used, and for UFC and SCM determination, chemiluminescent microparticle immunoassay was used. RESULTS The highest correlation coefficient characterised LSC and SCM (r = 0.72). Area under curve (AUC) for SCM and LSC in receiveroperating characteristic (ROC) for OCS was: 0.86 v. 0.74; for PCS: 0.83 v. 0.70; and for SCS: 0.74 v. 0.79. CONCLUSIONS Our findings suggest that LSC is more useful compared to SCM in diagnosing subclinical Cushings syndrome. Moreover,LSC seems to be a valuable diagnostic criterion to distinguish pseudo Cushings state. However, to obtain reliable cut-offs for LSC values,a larger group of hypercortisolic patients is needed.

Value of direct radionuclide cystography in diagnosing vesico-peritoneal fistulae

A 36-year-old female patient underwent a laparoscopic surgery to remove a uterine fibroid. The procedure failed to relieve the pelvic pain, although its nature changed. After a period of observation, the patient was re-admitted to hospital on suspicion of a vesico-uterine fistula to be differentiated with endometriosis. Diagnostic investigations - cystography, cystoscopy, computed tomography and magnetic resonance - did not reveal a fistula. Laparoscopy was performed, with a possible biopsy in order to eliminate endometriosis. The result was negative, but chronic progressive reactive/inflammatory lesions were noticed, possibly indicating the presence of a vesico-peritoneal fistula. Therefore, a direct radionuclide cystography was performed. The scintigraphic images single-photon emission computed tomography (SPECT/CT) showed a radioactive spot, indicative of a vesico-peritoneal fistula. The fistula was treated for three months by catheterisation of the urinary bladder. The follow-up SPECT-CT did not reveal any urine leakage from the bladder. The clinical symptoms disappeared as well.

Percentage of LFA-1+ and ICAM-1+ peripheral blood mononuclear cells in children and adolescents with type 1 diabetes does not distinguish patients with vascular complications.

There are only few studies evaluating lymphocytes activation in the diabetic vascular complications. ICAM-1/LFA-1 adhesion molecules not only participate in the lymphocyte T proliferation but also mediate leukocyte migration to the site of inflammation. We assess a relationship between the percentage of ICAM-1 and LFA-1 expressing PBMCs and the evolution of vascular complications in T1D in children and adolescents. The study was carried out on 60 children and adolescents with T1D (aged 9-20): (a) T1D lasting <5 years (n=20), (b) T1D lasting >5 years (n=20), without complications c) T1D lasting >5 years complicated with microalbuminuria, arterial hypertension, diabetic retinopathy (20 n). 20 healthy volunteers, age and sex matched constituted the control group. The expression of adhesion molecules was evaluated by using three-color flow cytometry. In children and adolescents with T1D <5 years, the percentage of ICAM-1+ and LFA-1+ PBMCs was decreased vs. controls (p<0.05 and p<0.001, respectively). Both in patients with T1D>5 years without vascular complications and in T1D with vascular disease the percentage of LFA-1+ T lymphocytes was significantly reduced in the peripheral blood (p<0.001 vs. healthy controls). In conclusion the percentage of LFA-1+ and ICAM-1+ PBMCs does not distinguish patients with vascular complications however decreased percentage of LFA-1+ PMBCs could serve as a nonspecific marker of the development of local inflammatory process in Type 1 diabetes.