Henryk Stepien

Evaluation of Interleukins, ACTH, Cortisol and Prolactin Concentrations in the Blood of Patients with Parkinson's Disease

Parkinsons disease (PD) is characterized by a markedly decreased number of nigrostriatal dopaminergic neurons. The pathogenesis of PD is still unknown; among other etiological factors, immunological abnormalities have been suggested. Recently, interleukin-2 (IL-2) has been hypothesized to be an endogenous cytokine that regulates striatal dopaminergic function. We examined the plasma concentrations of IL-1, IL-2, IL-6 and blood levels of ACTH, cortisol and prolactin of 21 patients with PD without any previous treatment. Age- and sex-matched subjects without any neurological or immune disorders were used as controls. Significantly higher serum concentrations of IL-2 in patients with PD were found. Treatment with antiparkinsonian drugs reduced IL-2 levels in these patients. Our results suggested a functional relationship between central dopaminergic and immune systems and a possible involvement of the latter in the pathogenesis of PD.

Effect of somatostatin on the proliferation of mouse spleen lymphocytes in vitro

The effect of somatostatin on the spontaneous proliferation of mouse spleen lymphocytes was investigated in vitro. The rate of 3H-thymidine incorporation was used as an index of lymphocyte proliferation. Somatostatin in a concentration of 10(-7) M enhanced the lymphocyte proliferation and abolished the antiproliferative effect of rat hypothalamic extract. Lower concentrations of somatostatin slightly decreased the lymphocyte DNA synthesis.

Matrix Metalloproteinases, Tissue Inhibitors of Matrix Metalloproteinases and Angiogenic Cytokines in Peripheral Blood of Patients with Thyroid Cancer

Stimulation of growth of endothelial cells from preexisting blood vessels, i.e., angiogenesis, is one of the essential elements necessary to create a permissive environment in which a tumor can grow. During angiogenesis, the matrix metalloproteinase (MMP) family of tissue enzymes contributes to normal (embriogenesis or wound repair) and pathologic tissue remodeling (chronic inflammation and tumor genesis). The proposed pathogenic roles of MMPs in cancer are tissue breakdown and remodeling during invasive tumor growth and tumor angiogenesis. Tissue inhibitors of metalloproteinases (TIMPs) form a complex with MMPs, which in turn inhibits active MMPs. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are unique among mediators of angiogenesis with synergistic effect, and both can also be secreted by thyroid cancer cells. The goal of the study was to evaluate the plasma blood concentration of VEGF, bFGF, MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, TIMP-1, and TIMP-2 in patients with cancer and in normal subjects. Twenty-two patients with thyroid cancers (papillary cancer, 11; partly papillary and partly follicular cancer, 3; anaplastic cancer, 5; medullary cancer, 3) and 16 healthy subjects (controls) were included in the study. VEGF, bFGF MMPs, and TIMPs were evaluated by enzyme-linked immunosorbent assay (ELISA). In patients with thyroid cancer, normal VEGF concentrations (74.29 +/- 13.38 vs. 84.85 +/- 21.71 pg/mL; p > 0.05) and increased bFGF (29.52 +/- 4.99 vs. 6.05 +/- 1.43 pg/mL; p < 0.001), MMP-2 (605.95 +/- 81.83 vs. 148.75 +/- 43.53 ng/mL; p < 0.001), TIMP-2 (114.19 +/- 6.62 vs. 60.75 +/- 9.18 ng/mL; p < 0.001), as well as lower MMP-1 (0.70 +/- 0.42 vs. 3.87 +/- 0.53; p < 0.001) levels have been noted. Increased plasma levels of MMP-3 and MMP-9 were also found in patients with medullary carcinoma. In conclusion, predominance of MMP-2 over TIMP-2 and TIMP-1 over MMP-1 as well as increased concentration of bFGF in peripheral blood are common features in patients with thyroid cancer.

Effects of two neuropeptides, somatoliberin (GRF) and corticoliberin (CRF), on human lymphocyte natural killer activity

The effect of corticoliberin (CRF) and somatoliberin (GRF) on the natural killer (NK) activity of human peripheral blood lymphocytes was investigated. NK activity was estimated by means of the radioactive chromium (51Cr) assay in which human leukemia K 562 cells serve as targets. Exposure of human lymphocytes (effector cells) to 10(-6) to 10(-10) M concentrations of CRF inhibited NK activity. NK activity was also suppressed by the same concentrations of GRF, but only when an effector:target cell ratio of 40:1 was used. In contrast, at effector:target ratios of 20:1 and 10:1, stimulatory effects of GRF were observed.

Inhibition of Rat Pituitary Tumor Cell Proliferation by Benzodiazepines in vitro

The effect of various benzodiazepines (peripheral-type receptor ligands: Ro 5-4864, PK 11135; central-type receptor ligands: clonazepam, Ro 15-1788, Ro 15-4513; mixed type: diazepam) on the proliferation of estrogen-induced rat pituitary prolactin-secreting tumor cells was studied in vitro. [3H]thymidine incorporation into DNA was used as an index of cell proliferation. It was found that tested peripheral- and mixed-type benzodiazepine receptor ligands significantly suppressed the pituitary cell proliferation in a dose-dependent manner (10(-4)-10(-8) M). The inhibitory effect of Ro 5-4864 was reversed by 5 x 10(-3) M calcium chloride. On the other hand, central-type benzodiazepine receptor ligands suppressed tumor cell proliferation only at the highest concentration studied (10(-4) M). Our results indicate that benzodiazepines might exert an antiproliferative action on pituitary tumor cell growth, and that this effect seems to be a calcium-dependent process.

Evaluation of the Levels of bFGF, VEGF, sICAM-1, and sVCAM-1 in Serum of Patients with Thyroid Cancer

Tumour growth and development depend on a complex cascade of angiogenic factors. The aim of the study is evaluation of the level of growth factors VEGF and bFGF, and adhesion molecules sICAM-1, sVCAM-1 in the serum of patients with papillary thyroid cancer. The study comprised 35 patients aged 21-68 years (mean age 46+/-14) who had papillary thyroid cancer diagnosed on the basis of thin needle aspiration biopsy, and were qualified for operative treatment. This group comprised 28 women and seven men. The control group was 26 healthy individuals. Serum concentrations of bFGF, VEGF, sICAM-1, and sVCAM-1 were evaluated by the enzyme-linked immunosorbent assay (ELISA) method. We have observed significantly higher mean concentrations of bFGF, VEGF, and sICAM-1 in the serum of patients with thyroid cancer compared with the control group. There was no significant difference between the sVCAM-1 concentrations of the thyroid cancer group and the control group.

Decreased 1-25 Dihydroxyvitamin D3 Concentration in Peripheral Blood Serum of Patients with Thyroid Cancer

BACKGROUND AND AIMS Vitamin D(3), in addition to its role in calcium homeostasis, has been recognized as playing a role in human cancer development. However, little is known about the association between vitamin D status and the development of thyroid cancer. This study aimed to investigate vitamin D metabolism by measuring 25(OH) D(3), 1-25 (OH)(2) D(3), PTH and calcium concentrations in the peripheral blood of patients with different forms of thyroid tumors. METHODS The 25-hydroxyvitamin D(3) ,1-25- dihydoxyvitamin D(3), PTH and calcium serum levels of 50 consecutive patients with epithelial thyroid cancer 27 cases of papillary cancers (PTC), 16 follicular cancers (FTC), and seven cases of anaplastic cancers (ATC) and 34 multinodular nontoxic goiter (MNG) were measured by specific immunoassay. The control group consisted of 26 healthy volunteers. RESULTS Our results revealed significantly lower 1-25 (OH)(2) D(3) concentration in the PTC group (22.67 pg/mL +/- 8.12; p <0.05), FTC group (16.09 pg/mL +/- 6.15; p <0.02) and ATC group (9.48 pg/mL +/- 5.18; p <0.02). Levels of 1-25 (OH)(2) D(3) varied by cancer stage and were also significantly different. A significant decrease in circulating 1-25 (OH)(2) D(3) concentration was found in patients with stage I (24.12 pg/mL +/- 6.77; p <0.05), stage II (16.93 pg/mL +/- 4.55; p <0.05), stage III (12.44 +/- 8.98; p <0.02) and in stage IVa (6.18 +/- 2.22; p <0.01). There were no significant differences when comparing serum levels of 25(OH) D(3), PTH or calcium concentrations among individuals with multinodular goiter, thyroid cancer and age- and sex-matched control volunteers. CONCLUSIONS Our study revealed that impaired vitamin D(3) metabolism may play an important role in thyroid follicular cell oncogenesis.

Hypothalamic-Pituitary-Thyroid Axis and the Immune System

The paper reviews data on bidirectional circuits between the hypothalamic-pituitary-thyroid (HPT) axis and the immune system. The effects of thyroliberin (TRH), thyrotropin (TSH) and of thyroid hormones (thyroxine and triiodothyronine) on the immune system, as well as the effects of mono- and lymphokines on the HPT axis are discussed.

Effects of diazepam on cell proliferation in cerebral cortex, anterior pituitary and thymus of developing rats

The effect of a single dose (5mg/kg b.w.) of diazepam on the cell proliferation in selected organs of 11-day old female Sprague-Dawley rats has been investigated. The stathmokinetic method (counting of mitoses after colchicine administration) has been used for evaluation of cell replication. The significant fall of the mitotic activity in the parietal cerebral cortex and the anterior pituitary gland was found. On the other hand, the increase of the mitotic activity in thymus was observed. The reported data, taken together with the previous observations from our and other laboratories, suggest the involvement of benzodiazepine receptor in the control of cell proliferation.

Growth Hormone Replacement Decreases Plasma Levels of Matrix Metalloproteinases (2 and 9) and Vascular Endothelial Growth Factor in Growth Hormone–Deficient Individuals

Background—Matrix metalloproteinases (MMP) are implicated in cardiovascular disease. Growth hormone (GH) deficiency is associated with increased cardiovascular mortality. We assessed whether GH replacement, in GH-deficient adults, has any effect on plasma levels of MMP-2 and MMP-9 and on vascular endothelial growth factor (VEGF), known to activate MMPs. Methods and Results—The study comprised 66 GH-deficient adults, 37.8±14.7 years of age (37 female). Plasma MMP-2 and MMP-9, VEGF, and insulin-like growth factor-1 (IGF-1) were measured at baseline (V1), at 12 months (V2), and at 24 months of GH treatment (V3). IGF-1 levels rose under GH replacement (mean±SD): V1, 151.6±91.9 μg/mL; V2, 270.2 7114.8 μg/mL; and V3, 266.2±109.8 (V1 versus V2; P <0.001: V2 versus V3; P =0.76). MMP-9 exhibited the most pronounced and sustained decline from 1248.0±651.1 ng/mL at V1, 949.2±457.7 ng/mL at V2, and 760.8±386.1 ng/mL at V3 (P <0.001 at all time points). A similar pattern was detected for VEGF levels: 358.5±209.0 pg/mL at V1, 310.6±225.7 pg/mL at V2 (P <0.001), and 283.7±202.7 pg/mL at V3 (V2 versus V3; P =0.005). MMP-2 demonstrated a significant decline initially from V1 to V2 (1134.4±217.8 ng/mL versus 1074.5±203.0 ng/mL, respectively; P =0.031), reaching a plateau at V3 (1072.3±220.2 ng/mL) (V2 versus V3; P =0.93). A negative relation existed between MMP-9 versus IGF-1 and MMP-2 versus IGF-1 (P <0.001 and P =0.007, respectively) as well as between VEGF and IGF-1 (P <0.001). Conclusions—These changes in MMPs and VEGF may contribute to the anticipated reduction in vascular mortality in hypopituitary adults receiving GH replacement.