Electronic Library of Scientific Literature
Volume 33 / No. 3 / 1999
A. Syrenicz, G. Kurzawski, A. Ciechanowicz
Department of Endocrinology, Hypertension and Metabolic Diseases;
Department of Genetics and Pathomorphology;
Department of Clinical Biochemistry, Pomeranian Medical Academy, 71-455 Szczecin, Poland
Objective. To evaluate the detection possibility of TSH receptor gene mutation
within the third cytoplasmic loop and the sixth transmembrane domain in the cytological
material obtained by means of fine needle biopsy of autonomous and non-autonomous nodules.
Methods. The study has been carried out in 16 women with goitre showing no clinical signs of hyperthyroidism. According to the thyroid scintigraphy and serum level of thyrotropin (TSH) the patients were divided into two groups: 1. 6 patients with autonomous nodules; 2. 10 patients with non-autonomous nodules. Genomic DNA has been isolated from the cytological material and the peripheral blood nuclear cells in order to confirm possible somatic character of TSH receptor gene mutations. DNA has been amplified in polymerase chain reaction (PCR) with the use of a specific pair of primers. Purified PCR products have been subjected to further automatic sequencing.
Results. Among 6 autonomous nodules tested one heterozygotic somatic mutation of adenine for cytosine at 1804 nucleotide of TSH receptor gene was detected. This mutation resulted in the change of threonine (codon ACC) at 632 position of TSH receptor protein for proline (codon CCC). Among the non-autonomous nodules one heterozygotic somatic mutation of adenine for cytosine at 1870 nucleotide of receptor TSH gene has been detected. From this mutation followed the change of lysine (codon AAG) at 624 position of the polypetide chain for glutamine (codon CAG) followed as a consequence.
Conclusions. We emphasize the validity of fine needle biopsy in the detection of somatic mutations in the TSH receptor gene. For the first time the somatic mutation in the TSH receptor gene in a non-autonomous nodule has been reported.
Key words: Thyroid nodules - TSH receptor - Somatic mutations - Fine needle biopsy
Endocrine regulations, Vol. 33, 95 - 101, 1999
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L. Starka, M. Hill, R. Hampl, I.T. Huhtaniemi
Institute of Endocrinology, 116 94 Prague, Czech Republic;
Department of Physiology, University of Turku, 20520 Turku, Finland
Objective. To evaluate the frequency of luteinizing hormone (LH) variant in males
and females in West region of the Czech Republic. This species of mutated LH concerns
borderline alterations in pituitary-gonadal function, including higher risk for the
development of more aggressive forms of prostate carcinoma in males.
Methods. The examined normal population consisted of randomly selected 82 males and 175 females (age range of 14 to 72 years). Variant LH was determined by immunofluorimetric method using two pairs of monoclonal antibodies, one of which detecting both wild-type (w+) and variant LH, while the other detecting only w+-LH.
Results. The carrier frequency of the variant LH allele in the population sample was 17.5 % (12.2 % in males and 20.6 % in females) which was within the range of the European prevalence.
Conclusion. The prevalence of the common variant of LH in the investigated region of West Bohemia was close to that of other Middle-European and North-European populations.
Key words: LH variant - Prevalence in Czech Republic - Immunofluorimetric
Endocrine regulations, Vol. 33, 103 - 108, 1999
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A. Ristic-Fira, M. Vujcic, M. Krstic-Demonacos, D. Kanazir
Laboratory for Molecular Biology and Endocrinology, Institute of Nuclear Sciences
”VINCA”, Belgrade, Yugoslavia;
University of Glasgow, Division of Biochemistry and Molecular Biology, Glasgow, Scotland, UK;
Serbian Academy of Sciences and Arts, Belgrade, Yugoslavia
Objective. To gain better insight into the role of glucocorticoids as modulators of
cell growth, as well as to investigate the presence and characteristics of glucocorticoid
receptors (GR) in mouse melanoma cells.
Methods. In two different B16 mouse melanoma cell clones (B16/F10 and B16/C3) the role of synthetic glucocorticoids (triamcinolone acetonide, TA) as cell growth modulators was investigated.
Results. The inhibitory effect of TA on B16/F10 cell growth after 8 days in culture was observed. The same hormonal treatment applied on B16/C3 melanoma cells also provoked changes in the cell growth. Dot blot analysis, using monoclonal antirodent glucocorticoid receptor antibodies showed the presence of receptor protein in both cell clones. The analysis of glucocorticoid receptors in B16/F10 and B16/C3 cell cytosol by Scatchard assay and ion-exchange chromatography on DEAE-Sephadex A-50 minicolumn indicated that the changes in melanoma cell growth may be mediated by glucocorticoid receptors and may relieve changes in the GR itself.
Conclusions. It was found that B16/C3 melanoma cells exhibited different growth pattern under TA treatment when compared to the results obtained with B16/F10 cells. Such differences may be mediated by glucocorticoid receptors.
Key words: Melanoma cells - Glucocorticoid receptor - Triamcinolone acetonide -
Endocrine regulations, Vol. 33, 109 - 115, 1999
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Przala J., T. Kaminski, G. Siawrys, S. Okrasa
Department of Animal Physiology, University of Agriculture and Technology, 10-718 Olsztyn-Kortowo 5, Poland
Objective. 1. To compare the release of beta-endorphin-like immunoreactivity
(beta-END-LI) by large and small luteal cells of the pig; 2. to test the effects of human
chorionic gonadotropin (hCG) either alone or combined with the cytokine TNFalpha on
beta-END-LI secretion by these cells.
Methods. Isolated large and small luteal cells on days 8-10 of the cycle (n=7) were incubated in M199 supplemented with hydrocortisone (40 ng/ml), transferrin (5 microg/ml), insulin (2 microg/ml), gentamicin (50 microg/ml), nystatin (240 U/ml), porcine low-density lipoproteins (LDL; 100 microg/ml), 1 % BSA and 2 x 10-5 M bacitracin, for 12 h at 37 °C and under the atmosphere of 95 % air and 5 % CO2. The cells were treated either with hCG (100 ng/ml) or TNFalpha (0.1, 1 and 3 nM) alone or with both agents together. beta-END-LI concentration in incubation media was measured by RIA.
Results. beta-END-LI secretion by large luteal cells was 15-fold greater than by small cells (666.38 ± 24.59 pg/ml/106 cells vs. 44.60 ± 2.53 pg/ml/106 cells). hCG enhanced beta-END-LI secretion by both large and small luteal cells. TNFalpha alone had no effect on beta-END-LI release by large and small luteal cells, but it abolished the stimulatory effect of hCG on beta-END-LI secretion by large luteal cells.
Conclusion. The results indicate that large luteal cells are a major source of beta-endorphin in the porcine corpus luteum, where its release may be affected by gonadotropins (possibly LH) and TNFalpha.
Key Words: Luteal cells - hCG - TNFalpha - beta-endorphin - Gilts
Endocrine regulations, Vol. 33, 117 - 123, 1999
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J. Galas, P. Epler, S. Stoklosowa
Laboratory of Animal Endocrinology and Tissue Culture, Institute of Zoology,
Jagiellonian University, 30-060 Krakow, Poland;
Department of Ichtyobiology & Fisheries, Agriculture University, 30-149 Krakow-Mydlniki, Poland
Objective. To study seasonal variation of steroid secretion by dispersed carp
ovarian follicular cells as influenced by carp pituitary homogenate (CPH), human chorionic
gonadotropin (hCG) or 17alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 betaOH-P).
Methods. Carp ovaries were collected each month throughout the year from January to December. The cells of ovarian follicles were dispersed by trypsin treatment and cultivated as monolayers. Control cultures were grown in medium M199 alone. Media for experimental cultures were supplemented either with CPH, hCG or (17alpha,20 betaOH-P). The direct effect of these three factors on progesterone (P4), androgens and estradiol (E2) secretion by isolated follicular cells in cell culture was assayed by appropriate RIA.
Results. In control cultures the higshest levels of P4, A and E2 appeared before spawning time and at the time of vitellogenesis. Under the influence of CPH the secretion of P4 increased in December. Significant increase of androgen secretion was seen in May, while that of E2 appeared in February, May, September and December. In December, February and November the secretion of E2 was suppressed by hCG. The most intense stimulation of steroidogenesis was observed after 17alpha,20 betaOH-P treatment. It increased the level of P4 at the time of its maximum in the control cultures; androgen level was elevated throughout the whole year; E2 secretion was stimulated only during the spawning time, while during vitellogenesis E2 level declined.
Conclusion. It was shown that steroid secretory pattern of follicular cells changed during the year and that the three factors used (pituitary homogenate, hCG and 17alpha,20 betaOH-P) exerted direct influence upon steroids secretion.
Key words: Carp ovarian cells - Progesterone - Estradiol - Androgens - hCG -
Seasonal variation - Tissue culture - Pituitary homogenate - 17alpha,20 betaOH-P
Endocrine regulations, Vol. 33, 125 - 133, 1999
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T. Mitsuma, N. Rhue, M. Kayama, Y. Mori, Y. Yokoi, K. Adachi, R. Ikai, A. Nakamura, A. Nakayashiki, T. Nogimori, J. Sakai, Y. Hirooka
Fourth Department of Internal Medicine,
First Department of Physiology and
Department of Laboratory Medicine, Aichi Medical University, Nagakute, Aichi, 480-1195 Japan;
Department of Internal Medicine, Konanshowa Hospital, Konan, Aichi
Objective. To investigate the organ distribuion of thyrotropin releasing hormone
receptor (TRHR) type 2 in rats by immunohistochemical method.
Methods. TRHR type 2 was identified immunohistochemically in the rat tissues using specific anti-TRHR antiserum raised in New Zealand white rabbits immunized with a conjugate of synthetic TRHR type 2 (5-23) with bovine serum albumin. Immunohistochemical analysis was performed by avidin-biotin complex method.
Results. TRHR type 2 immunoreactivity was visualized in the central nervous system, anterior pituitary, gastric mucosa, Auerbach’s and Meissner’s nervous branch of the stomach, small intestine and colon, retina amd testis. Significant stain was detected in neural perikarya, axons and dendrites. When using antiserum preincubated with synthetic TRHR type 2(5-23) or anterior pituitary homogenates, no significant stain of anterior pituitary was detected.
Conclusions. These findings suggest that TRHR type 2 is widely distributed and that the method used is valuable in studying the distribution of TRHR type 2 in rats.
Key words: TRH receptor type 2 - TRH - Immunohistochemistry - Organ distribution -
Endocrine regulations, Vol. 33, 135 - 139, 1999
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E. Ersoy, F. Taneri, E. Tekin, A. Poyraz, A. Cihan, A. Dursun, E. Onuk
Department of General Surgery and
Department of Pathology, Gazi University Medical Faculty, Besevler, Ankara, Turkey
Objective. In a double blind prospective clinical study to evaluate the diagnostic
potential of peroperative fine needle aspiration cytology as compared to peroperative
frozen section in thyroid surgery.
Methods. The diagnostic value of one hundred consecutive preoperative (FNA) and peroperative fine needle aspiration (p-FNA), frozen section (FS) and permanent section (PS) examination for thyroid nodules were studied prospectively in order to assess and compare the accuracy, sensitivity and specificity.
Results. Out of 100 patients PS showed 11 % of malignancies, while p-FNA showed 5 % and FS showed 6% of malignant cases with no false positive, but with 6 and 5 false negative results, respectively. Thus, as compared with FS, one false negative finding was obtained by p-FNA in a case of malignant tumor which could be definitely ascertained by frozen section technique. However, concerning the benign nodules no differences were found between p-FNA and FS.
Conclusions. Peroperative fine needle aspiration seems to be a useful method which can be properly performed because the nodule can be easily seen during the surgical procedure. However, further clinical observations of large numbers of patients are needed.
Key Words: Thyroid surgery - Peroperative FNA - Frozen sections - Diagnostic
Endocrine regulations, Vol. 33, 141 - 144, 1999
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