Identification of Recessively Inherited Genetic Variants Potentially Linked to Pancreatic Cancer Risk.

Although 21 pancreatic cancer susceptibility loci have been identified in individuals of European ancestry through genome-wide association studies (GWASs), much of the heritability of pancreatic cancer risk remains unidentified. A recessive genetic model could be a powerful tool for identifying additional risk variants. To discover recessively inherited pancreatic cancer risk loci, we performed a re-analysis of the largest pancreatic cancer GWAS, the Pancreatic Cancer Cohort Consortium (PanScan) and the Pancreatic Cancer Case-Control Consortium (PanC4), including 8,769 cases and 7,055 controls of European ancestry. Six single nucleotide polymorphisms (SNPs) showed associations with pancreatic cancer risk according to a recessive model of inheritance. We replicated these variants in 3,212 cases and 3,470 controls collected from the PANcreatic Disease ReseArch (PANDoRA) consortium. The results of the meta-analyses confirmed that rs4626538 (7q32.2), rs7008921 (8p23.2) and rs147904962 (17q21.31) showed specific recessive effects (p<10-5) compared with the additive effects (p>10-3), although none of the six SNPs reached the conventional threshold for genome-wide significance (p < 5×10-8). Additional bioinformatic analysis explored the functional annotations of the SNPs and indicated a possible relationship between rs36018702 and expression of the BCL2L11 and BUB1 genes, which are known to be involved in pancreatic biology. Our findings, while not conclusive, indicate the importance of considering non-additive genetic models when performing GWAS analysis. The SNPs associated with pancreatic cancer in this study could be used for further meta-analysis for recessive association of SNPs and pancreatic cancer risk and might be a useful addiction to improve the performance of polygenic risk scores.

Evidence for diagnosis of early chronic pancreatitis after three episodes of acute pancreatitis: a cross-sectional multicentre international study with experimental animal model.

Chronic pancreatitis (CP) is an end-stage disease with no specific therapy; therefore, an early diagnosis is of crucial importance. In this study, data from 1315 and 318 patients were analysed from acute pancreatitis (AP) and CP registries, respectively. The population from the AP registry was divided into AP (n = 983), recurrent AP (RAP, n = 270) and CP (n = 62) groups. The prevalence of CP in combination with AP, RAP2, RAP3, RAP4 and RAP5 + was 0%, 1%, 16%, 50% and 47%, respectively, suggesting that three or more episodes of AP is a strong risk factor for CP. Laboratory, imaging and clinical biomarkers highlighted that patients with RAP3 + do not show a significant difference between RAPs and CP. Data from CP registries showed 98% of patients had at least one AP and the average number of episodes was four. We mimicked the human RAPs in a mouse model and found that three or more episodes of AP cause early chronic-like morphological changes in the pancreas. We concluded that three or more attacks of AP with no morphological changes to the pancreas could be considered as early CP (ECP).The new diagnostic criteria for ECP allow the majority of CP patients to be diagnosed earlier. They can be used in hospitals with no additional costs in healthcare.

Higgs physics at the CLIC electron-positron linear collider.

The Compact Linear Collider (CLIC) is an option for a future [Formula: see text] collider operating at centre-of-mass energies up to [Formula: see text], providing sensitivity to a wide range of new physics phenomena and precision physics measurements at the energy frontier. This paper is the first comprehensive presentation of the Higgs physics reach of CLIC operating at three energy stages: [Formula: see text], 1.4 and [Formula: see text]. The initial stage of operation allows the study of Higgs boson production in Higgsstrahlung ([Formula: see text]) and [Formula: see text]-fusion ([Formula: see text]), resulting in precise measurements of the production cross sections, the Higgs total decay width [Formula: see text], and model-independent determinations of the Higgs couplings. Operation at [Formula: see text] provides high-statistics samples of Higgs bosons produced through [Formula: see text]-fusion, enabling tight constraints on the Higgs boson couplings. Studies of the rarer processes [Formula: see text] and [Formula: see text] allow measurements of the top Yukawa coupling and the Higgs boson self-coupling. This paper presents detailed studies of the precision achievable with Higgs measurements at CLIC and describes the interpretation of these measurements in a global fit.

Polymorphisms of the glucocorticoid receptor gene in Graves ophthalmopathy.

BACKGROUND/AIMS: Glucocorticoids have an important role in the regulation of the immune system, and alterations in glucocorticoid signaling may have an impact on the pathophysiology of autoimmune and inflammatory disorders. Because polymorphisms of the glucocorticoid receptor (GR) gene, including the N363S, ER22/23EK, A3669G and BclI variants were found to influence glucocorticoid signalling, we examined whether these polymorphisms could be associated with the development or clinical manifestations of Graves ophthalmopathy (GO). METHODS: The carrier and allelic frequencies of the N363S, ER22/23EK, A3669G, and BclI polymorphisms of the GR were determined in 95 Hungarian outpatients with GO and 160 healthy controls. RESULTS: No significant changes were found in carrier frequencies of the four polymorphisms between GO patients and healthy controls. However, when GO patients were divided into two subgroups (American Thyroid Association Committee, ATA I-II vs ATA III or greater), the frequency of the polymorphic BclI allele was significantly higher in patients with ATA I-II compared with those with ATA III or more (p = 0.009). CONCLUSION: The significant association between the frequency of the polymorphic BclI allele and ATA stage distribution suggests that this polymorphism of the GR gene may affect clinical manifestations of GO, presumably due to an increased signaling of endogenous glucocorticoids.

Soluble CD4 concentrations predict relapse of post-partum thyroiditis.

Post-partum thyroiditis (PPT) is a common autoimmune thyroid disorder which results in significant morbidity at a critical time of a woman's life. The presence of anti-thyroglobulin (anti-TG) and, more so, anti-thryroperoxidase (anti-TPO) antibodies in the first trimester of pregnancy has been reported to forecast subsequent PPT. Despite their predictive value, these tests lack in specificity. We have sought to find an alternative that is more specific and, ideally, which could be tested immediately proximate to the event. We have taken advantage of the high recurrence rate of PPT in subsequent pregnancies to perform a prospective study of serum soluble CD4 (sCD4) and CD8 (sCD8) levels in 22 pregnant women who had at least one previous episode of PPT. This group was matched with 21 pregnant women of comparable age with no evidence of thyroid disease. Both groups of women were sampled in each of the three trimesters of pregnancy, 1 month, 3 months and 6 months post-partum for sCD4, sCD8, thyroid function parameters and antibodies. Twelve of the 22 women with previous PPT had recurrent disease; they were more likely to be cigarette smokers and to have a family history of autoimmune disorders (p<0.05, for both) than those who did not. Half of these women had high anti-TG or anti-TPO each in the first trimester compared to none among those without recurrent PPT and 2/21 controls. Serum sCD8 levels showed no changes over the observation points among the two PPT patient subsets and were comparable to those of the controls. By contrast, serum sCD4 concentrations showed divergent changes in the group with recurrent PPT in the course of pregnancy and postpartum period compared to those without disease recurrence and controls: sCD4 failed to show the physiological fall in the third trimester of pregnancy [19.0+/-1.7 (+/-SD) U/ml vs 15.6+/-2.3 U/ml in controls, NS]. This trend was continued into the first month post-partum when sCD4 levels were clearly higher than in controls (22.1+/-2.6 U/ml compared to 17.9+/-1.9 U/ml in controls, p<0.001) and well before the episode of PPT. An sCD4 serum level outside the 95% reference range at 1 month post-partum (9/12 in recurrent PPT, 1/21 in controls) yields a relative risk of 6.9 (chi2=14.67, p<0.001) compared to 3.3 for first trimester thyroid antibody positivity (p=0.029). In summary, we describe a reliable test for forecasting PPT that can be obtained immediately proximate to the possible event. If our findings are verified in larger studies, the measurement of serum sCD4 concentration drawn in the first month post-partum may prove an ideal test for population screening for impending PPT.

The human thyrotropin receptor is highly mutable: a review of gain-of-function mutations.

OBJECTIVES: To find whether germline and somatic gain-of-function mutations of the thyrotropin receptor (TSHR) differ in location and/or mutational mechanisms, as well as to explore the degree to which these mutations are specific to TSHR compared with pituitary glycoprotein hormone receptors. METHODS: We examined the data on the TSHR website (www.unvi-leipzeig approximately innerre/TSH) supplemented with recent literature. Comparisons were also made with gain-of-function mutations of lutropin/choriogonadotropin (LH/CGR) and follicle-stimulating hormone receptors (FSHR). RESULTS: Some mutations (at residues 183, 505, 509 and 597) are exclusively germline, whereas mutations at 630 and 633 are characteristic of somatic mutations. Several residues located mainly in a mutation cluster region (619-639) are shared by both. Germline mutations are more likely to be transitions than transversions compared with somatic mutations. The lack of mutations involving deamination of CpG dinucleotides, a common mechanism for C-->T transitions, reflects the low CG prevalence in the mutable regions of TSHR. Comparison of the mutation sites with the equivalent positions in LH/CGR showed a significant difference (P<0.0001), whereas those in the mutation cluster region comprising the sixth transmembrane helix (TM6) and the adjoining third intracellular loop were concordant (P>0.90). We suggest that there is specific clustering of mutations in the juxtacytoplasmic end of TM6 in LH/CGR, a hydrophobic patch that is tightly packed with a face on TM5 whose sequences diverge from those of TSHR. CONCLUSIONS: TSHR exhibited frequent mutations outside the mutation cluster region. A role for a mutagenic environment created by the thyroid for other TSHR-specific codons cannot be discounted, nor can genetic factors, when accounting for the variation in the prevalence of TSHR-activating mutations worldwide.

[Current diagnostic method, prognosis estimation and therapy of papillary thyroid cancer: recommendations of the medical universities and the National Oncologic Institute of Budapest]. / A papillaris pajzsmirigy-carcinoma korszerü kivizsgálása, prognózisbecslése és kezelése: az orvostudományi egyetemek és az Országos Onkológiai Intézet ajánlása.

Physical examination, cervical ultrasonography (US) and aspiration cytology are the mainstays of the preoperative diagnostics of papillary thyroid carcinoma. For the staging of suspected malignant cases, cervical and mediastinal CT (MRI for inconclusive results) is indicated before any surgery. The end-result of primary treatment is assessed by total-body iodine scintigraphy and the serum human thyroglobulin (hTG) level. For long-term follow-up, physical examination and the serum hTG level are the most reliable tools (6-monthly), supplemented by cervical US and chest X-ray (yearly), and total-body iodine scintigraphy (2-yearly). If these furnish positive results, further examinations may be indicated. In suspected relapses of hTG non-producing and iodine non-accumulating papillary carcinomas, 201thallium chloride or 99mTc-sesta-MIBI (methoxy-isobutyl-isonitrile) scintigraphy, and positron emission tomography with 18fluoro-deoxyglucose or 11C-methionine may be of help. For estimation of the prognosis (cause-specific survival) of the patients, the MACIS score system of the Mayo Clinic is widely accepted, the patients being divided into low-risk and intermediate/high-risk categories. The recommended standard surgical intervention is near-total thyroidectomy (2-4 g residual glandular tissue left at the upper pole of the less-involved lobe), with a central cervical lymph node dissection for diagnostic purposes. In cases of lymph node dissemination, dissection (radical, modified radical, selective or microdissection) of any of the involved compartments (central, right or left cervical, or upper mediastinal) is indicated for therapeutic reasons, the method of which is depending on the extent of the metastatic involvement. Following adequate surgical intervention, no adjuvant radioiodine therapy is indicated for low-risk cases with a tumour of less than 1 cm diameter. For other low-risk or intermediate/high-risk patients, radioiodine ablation (R0N0M0) or a therapeutic radioiodine dosage (R2N1M1) is indicated. In cases at high-risk of local/regional relapse and in radioiodine non-accumulating tumorous cases, external radiotherapy may be applied. Thyroid hormone medication in a TSH suppressive dose is indicated during the first 5 postsurgical years: the goal is to achieve a TSH level below 0.1 (determined by a 3rd generation assay). If no relapse occurs or the case is a low-risk one, following the 5 years, it is enough to maintain the TSH level in a subnormal range (0.1-0.3).

Analysis of the genetic variability of the 1st (CCC/ACC, P52T) and the 10th exons (bp 1012-1704) of the TSH receptor gene in Graves' disease.

We determined the genetic variability of the 1st (CCC/ACC, P52T polymorphic variant) and 10th exons (bp 1012-1704) of the TSH receptor (TSHR) gene in Graves' disease. A total of 101 Graves' patients and 163 control subjects were screened. The A253 mutant allele was carried by nine patients with Graves' disease (8.91%) and 13 control subjects (7.98%) in heterozygous genotype. No significant difference in the frequency of the mutant allele was found between Graves' patients and control subjects. These results provide evidence that the A253 polymorphism has no genetic relevance in Graves' disease. Moreover, the DNA nucleotide sequence of 693 bp of the 10th exon (bp 1012-1704) of the TSHR gene was determined in 15 Graves' patients. Six patients were homozygous for the wild-type allele and nine were heterozygous for the mutant allele at the 253rd nucleotide of the first exon. No polymorphism was found in the DNA sequences obtained from leukocytes of Graves' patients, similarly to the sequences obtained from the nine control subjects. None of the nine patients carrying the A253 polymorphism in the 1st exon of the TSHR had polymorphism in the examined part of the 10th exon, including two additional patients whose thyroid tissue was directly analysed. In all likelihood, the polymorphisms of the examined regions of either the 1st or the 10th exon of the THSR gene do not contribute to the genetic susceptibility to Graves' disease.

Inhibitory effect of pentoxifylline on HLA-DR expression and glycosaminoglycan synthesis by retrobulbar fibroblasts.

OBJECTIVE: Glycosaminoglycan (GAG) production by retro-ocular fibroblasts (REF) is increased in patients with thyroid-associated ophthalmopathy (TAO). Various cytokines stimulate REFs to proliferate and elaborate GAG, free oxygen radicals as well as induce HLA-DR expression on these cells. Pentoxifyllin (Ptx) regulates the production of several cytokines including tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and, interferon gamma (IFN-gamma). We wished in this study to determine whether Ptx modified the spontaneous and cytokine-induced GAG synthesis by REF and IFN-gamma induced HLA-DR expression. DESIGN: REF derived from extraocular muscles of healthy subjects were cultured without and with cytokines (IFN-gamma, TNF alpha and IL-1) and the effect of Ptx on the production of GAG by REF and HLA-DR expression was determined. MEASUREMENTS: Glycosaminoglycan was measured by incorporation of (3H) glycosamine into GAG. HLA-DR expression was analyzed by fluorescence activated cell sorter. RESULTS: Both spontaneous and cytokine induced GAG synthesis by REF was inhibited by Ptx (100, 500 and 1000 mg/l, respectively). IFN-gamma (50, 100 and 500 U/ml) induced a dose-dependent increase in the expression of HLA-DR molecules by REF. Ptx, which was not toxic to REF, inhibited HLA-DR expression on those cells dose-dependently. CONCLUSIONS: Our in vitro results suggest that Ptx reduces cytokine-induced GAG production and HLA-DR expression by REF. It thus has potential as a therapeutic agent which regulates the function of lymphocytes infiltrating the retro-orbital tissues, and which are instrumental in TAO.

Effect of anti-thyroid peroxidase (TPO) antibodies on TPO activity measured by chemiluminescence assay.

A chemiluminescence method was developed to measure thyroid peroxidase (TPO) activity and the inhibitory effect of anti-TPO antibodies in purified porcine TPO. The TPO preparation was characterized kinetically and controlled by Western-blotting technique. The chemiluminescence method proved to be reproducible and much more sensitive than the widely used guaiacol method, being able to detect TPO concentrations of 2.21 x 10(-5) g/L vs 6.63 x 10(-2) g/L with the latter. Otherwise, the determinations with the two methods correlated well (r = 0.76). Investigating the effect of IgGs from 23 hypothyroid patients on measured TPO activity, we detected inhibition in 19 cases with the chemiluminescence technique (15 with the guaiacol method). Anti-TPO antibodies showed competitive inhibition of TPO activity with respect to the substrate guaiacol. In both systems, the inhibition is present in the IgG F(ab')2 fragment. We conclude that the high sensitivity of chemiluminescence detection allows routine determination of the inhibition of TPO activity by anti-TPO antibodies.

[Polymorphism of the 52 triplet gene (nucleotide 253) of the TSH receptor in Basedow-Graves patients and in healthy controls]. / A TSH-receptor gén 52. tripletjének (253. nukleotid) polimorfizmusa Basedow-Graves-kóros betegekben és egészséges egyénekben.

It has been analysed the polymorphism of the first exon of TSH receptor gene, at the first nucleotide of the triplet 52 of the genomic DNA (CCC/ACC, Pro/Thr) in Graves' disease and control population and was not found connection between the genetic background, clinical picture and some immunological parameters. Genomic DNA was obtained from formalin-fixed, paraffin embedded thyroid tissue of patients with Graves' disease underwent subtotal thyroidectomy and from peripheral blood leukocytes. The first exon and small part of the first intron of the TSH-R gene was amplified by PCR. The motif was amplified by the primers and primers includes restriction site of the Tth 111 I restriction enzyme and mutant form of the examined nucleotide. Genomic DNA having only wild allele was detected at 189 bp, while mutant allele was digested a 167 and a 22 bp fragments. Three persons (two female and one male) of the 32 patients with Graves' disease had mutation A253 in heterozygotic form: The patient with mutation A253 had no any symptoms of Graves' ophthalmopathy. No correlation was found between mutation A253 and the levels of anti-TSH-R-, anti TG, anti-TPO and anti-eye muscle antibodies. Surprisingly, it was found mutation of heterozygous genotype in two control individuals (one female and one male) of 14 without any symptoms of thyroid disease and negative laboratory findings. Finally, it has not been found the association of the 253 nucleotide of the codon 52 polymorphism with Graves' disease and ophthalmopathy and the allele A253 has no pathogenetic relevance in Graves' disease. On the other hand, it cannot be excluded that other mutation or sequence polymorphism in the remainder of TSH-R extracellular domain might be important in autoimmune mechanism of Graves' disease.

Beneficial effect of pentoxifylline on thyroid associated ophthalmopathy (TAO)*: a pilot study.

We have previously found that pentoxifylline (Ptx) inhibited cytokine induced HLA-DR expression and glycosaminoglycan (GAG) synthesis by retroorbital fibroblasts. We have now tested the clinical efficacy of Ptx in treating TAO. Ten patients with moderately severe ophthalmopathy were selected for study. All patients were euthyroid before and during the 12 weeks of the Ptx therapy. Serum GAG, TNF-alpha, anti-TSH-receptor, anti-eye muscle, anti-thyroglobulin and anti-thyroid peroxidase antibodies were determined sequentially. At the end of 12 weeks eight of the ten patients showed improvement in soft tissue but not in proptosis or extraocular muscle involvement. At baseline the levels of GAG (5.2+/-0.92 mg/dl v.s. 0.7+/-0.14 mg/dl, p<0.001) and TNF-alpha (33.6+/-6.6 pg/ml v.s. 5.4+/-1.3 pg/ml, p<0.001) were increased in patients compared to controls. They gradually decreased in the eight patients who responded to Ptx: after 4, 8 and 12 weeks of therapy serum GAG was 3.4+/-0.42 mg/dl, 2.5+/-0.77 mg/dl (p<0.01) and 1.1+/-0.2 mg/dl (p<0.001), respectively and serum TNF-alpha was 20.9+/-4.8 pg/ml, 14.9+/-2.2 pg/ml (p<0.05) and 9.7+/-1.8 pg/ml (p<0.01), respectively. Serum GAG and TNF alpha did not fall in the two patients who did not respond. The titre of anti-eye muscle antibodies but not anti-thyroid antibodies were lower at 12 weeks. Ptx has a beneficial effect on inflammatory symptoms of TAO and associated laboratory parameters in the majority of patients.

Effect of TSH and anti-TSH receptor antibodies on the plasma membrane potential of polymorphonuclear granulocytes.

Effects of thyrotropin hormone (TSH) and anti-TSH receptor antibodies on the plasma membrane potential of polymorphonuclear granulocytes (PMN) were analyzed by means of flow cytometry. Both TSH and the autoantibody caused a rapid, dose-dependent hyperpolarization of the plasma membrane of PMNs. TSH was also able to mask (revert) the depolarizing effect of a chemotactic peptide, fMLP, on PMNs. No detectable rise in the cytosolic free calcium level accompanied the observed hyperpolarization. Quinine, a blocker of Ca(2+)-activated and voltage-gated K+ channels did not affect the hyperpolarization by TSH and antibodies. Decreasing the [K+] gradient across the plasma membrane by valinomycin, however, blocked the hyperpolarizing effect. Peptide362-376 (derived from the extracellular domain of TSH receptor) also blocked the hyperpolarization induced by both TSH and anti-TSHR antibodies. These data suggest that the observed hyperpolarization is a specific, receptor-mediated early signal during interaction of PMNs with TSH or anti-TSHR antibodies.

High circulating IL-6 level in Graves' ophthalmopathy.

IL-6 is a paracrine and autocrine cytokine, which acts in the regulation of immunological and inflammatory processes. Its production can be observed in different cell types, as well as thyrocytes. The purpose of the study was to examine the serum IL-6 levels between the patients with Graves' disease (N = 47) and without (N = 29) ophthalmopathy in respect of the presence of inflammatory eye signs and thyroiditis, thyroid function and radioiodine or medical treatments. The serum IL-6 levels were greater (P < 0.025) in the patients with ophthalmopathy (440 +/- 32.4 pg/ml) than in those without eye disease (81.6 +/- 25.2 pg/ml). An elevated serum IL-6 levels could be detected in 22 out of 47 patients with ophthalmopathy with longer manifestation of thyroid disease than one year in comparison with those who had shorter (694 +/- 35.3 pg/ml vs 215.8 +/- 27.9 pg/ml, P < 0.05). The increase showed a strong association with the inflammatory signs of eye disease in the patients with Graves' hyperthyroidism compared with those without ophthalmopathy (513.3 +/- 33.7 pg/ml vs 96.9 +/- 12.1 pg/ml, P < 0.025). Euthyroid function and the presence of thyroiditis did not influence the serum IL-6 levels. Radioiodine and medical treatments did not lead to a remarkable decrease in the serum IL-6 levels. The results supported that IL-6 cytokine may be an important factor in the inflammatory events of Graves' ophthalmopathy.

Inhibitory effect of pentoxifylline on HLA-DR expression and glycosaminoglycan synthesis of retrobulbar fibroblasts induced by interferon gamma.

Glycosaminoglycan (GAG) accumulation produced by retroocular fibroblasts (REF) has been observed in patients with thyroid-associated ophthalmopathy (TAO). Various cytokines are able to express HLA-DR molecules and stimulate the REF to proliferate GAG and free oxygen radicals. Pentoxifylline (Ptx) is known to have complex immunomodulatory effects on production of cytokines including interferon gamma (IFN-gamma). Ptx has been assumed to inhibit the cytokine-induced production of GAG and HLA-DR expression. We wished to determine whether Ptx has an effect on the IFN-gamma induced HLA-DR expression and influences the spontaneous and cytokine-induced GAG synthesis of REF. REF derived from extraocular muscles of healthy subjects were cultured without and with IFN-gamma. The effect of Ptx on expression of HLA-DR molecules and the production of GAG by REF was determined. Glycosaminoglycan was measured by incorporation of (3H)glycosamine into GAG. HLA-DR expression was analysed by fluorescence activated cell sorter. IFN-gamma (50, 100 and 500 U/ml) induced an increase in expression of HLA-DR molecules of REF. Ptx was proved not to be toxic for cultured cells. This drug was able to dose-dependently inhibit HLA-DR expression of REF. Both spontaneous and IFN-gamma-induced GAG synthesis of REF was inhibited by Ptx (100, 500 and 1000 mg/l, respectively). Due to in vitro inhibitory effects, Ptx is potentially able to modify the antigen presentation and the GAG synthesis by REF and it might be a useful therapeutical drug in the treatment of TAO.

[Bone metabolism markers in patients with Basedow-Graves disease]. / A csontanyagcsere markereinek vizsgálata Basedow-Graves-kóros betegekben.

Collagen type I is the main collagen type found in bones. Carboxyterminal propeptide, deriving and cleaved from procollagen type I (PICP) during collagen synthesis, is delivered into the blood, where it might represent an useful marker of bone formation similarly to osteocalcin. PICP, osteocalcin, alkaline phosphatase, serum and urinary calcium excretion were measured in 58 premenopausal females affected by Graves' disease and also 28 of them after attainment of euthyroidism by methimazole treatment to study these biochemical indices of bone remodelling before and after treatment. Before therapy PICP (mean +/- S.D.: 244.2 +/- 112.3 vs. 136.8 +/- 32.4 micrograms/l), osteocalcin (mean +/- S.D.: 17.8 +/- 6.7 vs. 7.5 +/- 2.7 micrograms/l) and other markers were significantly (p < 0.05) higher than sex and age matched controls (n = 24). Treatment induced a significant decrease of PICP, alkaline phosphatase, calcaemia and calciuria compared to pretreatment values, while osteocalcin did not significantly differ (mean +/- S. D.: 17.8 +/- 6.7 vs. 14.7 +/- 8.7 micrograms/l). These data suggest that hyperthyroidism due to Graves' disease causes an increase of serum levels of these markers, but further studies are necessary to asses the differences between PICP and osteocalcin as markers of osteoblast activity in hyperthyroidism.

[Immunomodulatory effect of pentoxifylline in Graves ophthalmopathy]. / A pentoxifyllin immunmoduláns hatása Graves-ophthalmopathiában.

In the pathomechanism of the thyroid associated ophthalmopathy (TAO) the inflammatory cytokines produced by infiltrating lymphocytes of the retroorbital tissues are involved. The activated lymphocytes have been shown to secrete a number of cytokines including tumour necrosis factor-alpha, interleukin-1 and interferon-gamma. The widely used immunosuppressive therapies have potential serious side effects. The pentoxifylline (Ptx) is known to have effect on production of cytokines. The aim of this study was to investigate the effect of Ptx on expression of HLA-DR molecules and production of glycosaminoglycan of human retroorbital tissue cultures and potential efficacy in patients with TAO. It was found that pentoxifylline (Ptx) was able to inhibit significantly the HLA-DR expression and glycosaminoglycan synthesis induced by inflammatory cytokines including TNF-alpha, IFN-gamma and IL-1. Ten patients with untreated moderate severe ophthalmopathy (8 female and 2 male) were excluded from steroid treatment due diabetes mellitus and psychiatric disease. Classification of eye changes was made by NOSPECS categories and total eye score. All patients were euthyroid during the study and was no remarkable difference in thyroid function and eye symptoms. Before and during Ptx therapy the laboratory parameters were also determined including glycosaminoglycan. TNF-alpha, anti-TSH-receptor, anti-eye muscle, anti-thyroglobulin and anti-thyroid peroxidase antibodies in the patients'sera. It was found a remarkable improvement in the eye symptoms in eight of ten patients. The levels of glycosaminoglycan (uronic acid) and TNF-alpha gradually decreased in eight patients who considered to be responders. The levels of uronic acid in plasma of the responders were found to be significantly lower after Ptx treatment. Before Ptx therapy the TNF-alpha in the sera was not different remarkably in non-responders and responders. After 4 weeks Ptx treatment the TNF-alpha decreased significantly in responders compared to non-responders (20.9 +/- 4.8 pg/ml v. s. 28.3 +/- 6.1 pg/ml) (p < 0.01). The titre of anti-eye muscle antibodies were found to be lower at the end of observation, however, the anti-thyroid antibodies were not changed remarkably. It was concluded that Ptx in the majority of patients (8/10) has a beneficial effect on inflammatory symptoms of TAO and laboratory parameters and suggested to use as an additive therapy, however, further comparative studies are required for final evaluation of Ptx in the treatment of TAO.

Detectable serum IgE levels in Graves' ophthalmopathy.

Graves' ophthalmopathy is an organ-specific autoimmune disorder in which the target organs are infiltrated by T lymphocytes and polymorphonuclear neutrophils that release immunoregulatory cytokines in the thyroid and retrobulbar tissues. Th2-type cytokines (IL-4, IL-5, IL-6) support the inflammatory symptoms and immunoglobulin production, e.g. IgE isotype. IgE is thought to play a part not only in allergy but also in the normal immune responses, antigen processing and presentation. Since studies report IgE accumulation around the extraocular muscles in ophthalmopathy and a correlation between the total IgE levels and the severity of ophthalmopathy in Graves' disease, we measured the total IgE levels in 56 patients with Graves' disease (associated with ophthalmopathy in 47 patients) and in 42 healthy subjects as controls to determine if IgE plays a part in the autoimmune or the local inflammatory processes. For detection we used the Enzymun-Test IgE, which was a two-step ELISA sandwich assay. Elevated circulating IgE levels could be demonstrated in non-treated hyperthyroid Graves' patients in comparison with the controls (51.02 +/- 113.16 IU/ml vs 34.95 +/- 54.91 IU/ml, P < 0.01). The IgE levels were higher in patients with active inflammatory eye signs (63.65 +/- 130.41 IU/ml) than in controls (P < 0.007). The anti-thyroid drug and glucocorticoid management decreased the total IgE levels, and the difference was significant after the treatment compared with the values at the beginning of the therapy in the Graves' patients without ophthalmopathy (17.34 +/- 6.34 IU/ml vs 13.01 +/- 9.86 IU/ml, P < 0.03). In conclusion, since the results suggested that IgE plays a role in the inflammatory eye signs in Graves' ophthalmopathy, we propose administering antihistamines for medical management.