Electronic Library of Scientific Literature 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
estimation
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 -
Cell growth
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 -
rat
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
validity
Endocrine regulations, Vol. 33, 141 - 144, 1999
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