SEMA3A and IGSF10 Are Novel Contributors to Combined Pituitary Hormone Deficiency (CPHD).

Background: The mutation frequencies of pituitary transcription factors genes in patients with combined pituitary hormone deficiencies (CPHD) vary substantially between populations. However, apart from PROP1 the mutation rate of other genes is low and for almost half of the patients with CPHD the routine sequencing of known genes is unsuccessful in the identification of genetic causes. Methods: A cohort of 66 sporadic and nine familial CPHD cases (80 patients in total) were subjected to initial testing of the genes PROP1, POU1F1, LHX3, LHX4, and HESX1 using a targeted gene panel and MLPA. In patients who tested negative, a whole exome sequencing approach was employed. Results: In nine of the familial cases and 32 of the sporadic patients mutations in the PROP1 gene were found (the common pathogenic variants included c.301_302delAG and c.150delA). Mutations were also found in genes so far not related directly to CPHD. A unique homozygous and clinically relevant variant was identified in the SEMA3A gene, which may contribute to neural development and his phenotypic spectrum including short stature and isolated hypogonadotropic hypogonadism (IHH). Another pathogenic variant p.A1672T was found in the IGSF10 gene reported to be responsible for delayed puberty and neuronal migration during embryogenesis. Several suspected novel but predicted benign variants were also identified for the CHD7, WDR11 and FGF17 genes. Conclusion: Although PROP1 defects account for a majority of CPHD patients, identification of rare, less frequent variants constitutes a big challenge. Multiple genetic factors responsible for CPHD are still awaiting discovery and therefore the usage of efficient genomic tools (i.e., whole exome sequencing) will further broaden our knowledge regarding pituitary development and function.

Molecular Detection and Incidence of Y Chromosomal Material in Patients with Turner Syndrome.

The presence of a Y chromosome in patients with Turner syndrome (TS) is a risk factor for the development of gonadal tumor and/or virilization. With conventional cytogenetic analysis, some cells containing a Y chromosome can be missed. The aim of this study was to determine the presence and incidence of Y chromosome-derived material in TS patients using PCR and the markers SRY, DYZ1, DYZ3, DYS132, ZFY, and TSPY. Fifty-five TS patients (aged 5.5-26.75 years) were analyzed. A total of 17/55 (30.9%) were Y-positive, but only 7/17 had a Y chromosome in their karyotype and underwent gonadectomy. In 2 of these patients (28.6%), histopathologic examination revealed gonadoblastoma and dysgerminoma, respectively. In 8 patients in the studied group (8/55; 14.5%), the TSPY gene was detected, and the SRY gene (or a fragment) was identified in 9(3)/55 patients. No coding region mutations were observed in these SRY-positive patients. In conclusion, we have shown a high prevalence of Y chromosomal material in TS. Y markers were also observed in patients who had no Y chromosome in their karyotype, and PCR is very precise in detecting the presence of genetic material from the Y chromosome. Further follow-up of these Y-positive TS patients is mandatory.

Pituitary Microsomal Autoantibodies in Patients with Childhood-Onset Combined Pituitary Hormone Deficiency: an Antigen Identification Attempt.

The role of autoimmunization in the pathogenesis of pituitary disorders is poorly understood. The presence of pituitary autoantibodies (APA) has been detected in various pituitary disorders. Their role, however, remains elusive. Childhood-onset combined pituitary hormone deficiency (CPHD) may be caused by environmental or genetic factors. In some of patients, causes of the disease remain unclear and contributions of autoimmune processes have been postulated. The aim of this study was to identify the microsomes-derived pituitary antigens (MPA) as potential immunogenic autoantigens in patients with hypopituitarism, therefore 62 CPHD patients, 100 healthy controls and five autoimmune polyglandular syndrome type II (APS II) patients were included in the study. The clinical evaluation included hormonal tests and magnetic resonance imaging of the pituitary. The sources of MPA were pituitary glands taken from autopsies. Isolated MPA were then separated on SDS-PAGE gel and incubated with sera obtained from patients and controls. Microsomal APA were detected using Western blot and radioimmunological method. In all CPHD and APS II patients and in 9 % individuals from control group marked immunoreactivity was detected against MPA. Antibodies showed high affinity to 67, 60, 50 and 36 kDa MPAs. Since the identified autoantigens were of unknown nature, an in silico exploration of UniProt database was applied and indicated their possible relationship with chaperones, golgins and already known autoantigens like GAD67. Reactivity against MPA indicates that these proteins certainly play a role in the processes undergoing within pituitary of CPHD patients. The identification and further detailed studies on their role in the pathogenesis of CPHD should be continued.

The clinical role of serum concentrations of selected cytokines: IL-1ß, TNF-α and IL-6 in diagnosis of autoimmune thyroid disease (AITD) in children.

Chronic autoimmune thyroiditis (cAIT) leads to hypothyroidism due to T cell-mediated cytotoxicity in most cases. By contrast, Graves' disease (GD) with thyrotropin receptor stimulatory autoantibodies cause hyperthyroidism. Cytokines play a crucial role in modulating immune response in both disorders. The aim of study was to evaluate the concentrations of cytokines: IL-1ß, TNF-α and IL-6 in these two opposite clinical and hormonal thyroid diseases. The study group consisted of 64 children, 44 newly diagnosed, untreated children with cAIT (n = 22; with hypothyroidism) and GD (n = 22; hyperthyroidism), and the control group of 20 healthy children. Cytokine concentrations were evaluated using the ELISA technique. The studied groups of children did not differ significantly in concentrations of IL-6 (p = 0.48) and TNF-α (p = 0.067). In children with hypothyroidism, we found significantly higher concentrations of IL-1ß (median 2.16 pg/ml, IQR 0.87) compared to hyperthyroidism (median 1.39 pg/l, IQR 1.27) (p < 0.01) and the control group (median 1.88 pg/ml, IQR 1.04) (p < 0.05). The results of ROC curve analysis demonstrated the usefulness of IL-1ß (AUC = 0.77, p = 0.003) and TNF-α (AUC = 0.691, p = 0.034) as diagnostic parameters in cAIT which enable discrimination of children with autoimmune thyroid disease from healthy individuals. Concentrations of these markers are increased in autoimmune hypothyroidism. We found no significant sex differences in the tested parameters. In conclusion, IL-1ß and TNF-α may be considered as markers of hypothyroidism, and could efficiently discriminate between healthy and autoimmune hypothyroid children. Significantly higher concentrations of IL-1ß in children with hypothyroidism may be used to distinguish children with cAIT from GD patients.

[Estimation of growth hormone secretion during sleep as a screening test in the diagnosis of GH deficiency]. / Ocena wydzielania hormonu wzrostu po zasnieciu jako badanie przesiewowe w diagnostyce somatotropinowej niedoczynnosci przysadki.

BACKGROUND: Because of many disadvantages of growth hormone (GH) stimulation tests the diagnosis of growth hormone deficiency (GHD) is still problematic for the clinician. THE AIM of the study was the estimation of diagnostic usefulness of the GH provocation tests and basal IGF-I concentration measurement in the diagnosis of GHD. MATERIAL AND METHODS: The study group consisted of 180 children with short stature diagnosed in the Department of Pediatric Endocrinology and Diabetes in the years 1998-2003. Pharmacological stimulation test with insulin, clonidine, glucagon and L-dopa were used. GH concentration in physiological test after the onset of sleep was also measured. In 60 patients IGF-I concentration was estimated. For statistical analysis Spearman test was used. RESULTS: The highest mean GH concentrations (19.18 microl U/ml) and GH peaks (26.39 microl U/ml) were observed in sleep test. IGF-I concentration was correlated with GH concentration levels in physiological test after onset of sleep (r=0.4; p<0.05). CONCLUSIONS: Sleep is the strongest stimulatory agent for GH secretion. The estimation of GH secretion after the onset of sleep can be used as a screening test in the diagnosis of GHD. Because of many diagnostic problems in estimation of pituitary function the auxologic parameters should be considered as the most important part in the diagnostic work-up of children with short stature due to GHD.

The effects of growth hormone treatment in patients with somatotropin deficiency during their developmental age.

INTRODUCTION: In Poland treatment with growth hormone of adolescent patients dates back to 1964. Till 1993 the therapy was conducted in an interrupted manner, depending on the periodic availability of the drug. The data form such forms of therapy suggested that the end height within 3rd centile was achieved only by a portion of treated patients. Since 1995 the growth hormone is used in continuous therapy, which allows to sum up the effects of the therapy, including the growth rate and end height. MATERIAL AND METHODS: A total of 117 children and adolescent of both sexes, aged 4.6 to 18.1 years, with diagnosed somatotropic or multihormonal pituitary insufficiency were included in the study. All of them were treated with growth hormone and had an analysis of growth rate and end height. RESULTS: In the first 6 months of growth hormone treatment the growth rate achieved 10.4 cm/year in boys and 10.0 in girls and showed no correlation with maturation status. In the second half of the year the growth rate declined slightly. During the remainder of the therapy the growth rate markedly declined, and this effect was most notable in girls. In 93% of patients after the end of therapy the final height was no different than the expected height. CONCLUSIONS: 1. The growth rate in first half a year of the treatment was 3 times higher than before the beginning of therapy. 2. In the second half of the first year the growth rate slightly declined. 3. In following years the growth rate declined notably. 4. The final achieved height in most of the patients does not differ from the prognosed height.

[Tobacco smoking in patients with hyperthyroidism]. / Nikotynizm u chorych z nadczynnoscia tarczycy.

The objective of this study was to determine the impact of cigarette smoking in hyperthyroidism patients. The study group included patients with Graves-Basedow disease (GB): n = 317 (32.9%), patients with Graves' ophthalmopathy (GO): n = 108 (11.2%), patients with toxic nodular goiter (TNG) n = 511 (53%) and 28 (2.9%) patients with toxic adenoma (TA). Evaluation of tabacco smoking and exposure to environmental tobacco smoke (ETS) was performed on the base of questionnaire acording to Fagerström test. The comparison of GO and GB indicate increased tabacco smoking in GO (OR = 0.36; 95%CI: 0.22-0.59). The frequencies of tacacco smoking displayed significantly increased in GB and TNG patients (OR=0.39; 95%CI: 0.28-0.56). Further analys of patients with GO and TA presented statistcal significance in group of smoking and non-smoking (OR=0.26; 95%CI: 0,073-0,95). In other cases we didn't found a significant influence of tobacco smoking on thyroid disease. The effect of smoking was more pronounced in Graves' patients (particulary in the patients with GO) than in other thyroid patients. Smoking among patients with thyroid disease (GO and GB disease) is associated with developing of anxiety and fright, depression and problems with social relations sphere.

[Urinary cotinine in patients with hyperthyroidism]. / Kotynina w moczu u pacjentów z nadczynnoscia tarczycy.

The aim of our study was an analysis of cotynine, the main nicotine metabolite in the urine among hyperthyroid patients. The study group included 39 females and 4 males. The mean age was 35.59+/-14.22 yrs. (range: 18-73 yrs.; median: 32 yrs) among hyperthyroid patients suffering from: Graves-Basedow disease (GB), Graves' Ophtalmopathy (GO) and toxic nodular goiter (TNG). To evaluate the nicotine smoking intensity and ETS Environmental Tobacco Smoke, the urine analysis of cotynine level were performed. According to the statistical analysis using the Mann-Whitney test, the statistically significant difference between the level of cotynine among smokers suffering from GO and Graves-Basedow disease was revealed (p = 0.03). Similar results were obtained among the GO and TNG (p=0.02) using t-Student test with Welsch correction. To compare, there was no stastistically significant difference between the GB and TNG series (p=0.4). In the group of smoking patients with GO we found out incresed level of urine cotinine than in smoking patients with GB and TNG. We didn't found differences between GB and TNG in depends on an urine analysis of cotynine level.

[High level of hsCRP in patients with Graves' ophthalmopathy--preliminary study]. / Wysokie stezenie hsCRP u pacjentów palacych z oftalmopatia w przebiegu choroby Gravesa--doniesienie wstepne.

UNLABELLED: C-Reactive Protein (CRP) levels are used to indicate the presence of an inflammatory process that may be caused by bacterial infection or pathological processes. The objective of this study was to compare the levels of CRP and hsCRP (high sensitivity CRP) in smoking and non smoking patients with Graves' Ophthalmopathy (GO). Study included 59 hyperthyroid patients (age > 18 years, 50 females, 9 males, mean +/- SD: age 34 +/- 16) with varying degrees of severity of GO (classification according to NOSPECS) with hyperthyroidism prepared to radioiodine therapy. Smoking status was determined by Fagerström test and Brinkman's index. CONCLUSIONS: 1. Smoking additionally stimulates inflammatory processes in the patients with Graves' Ophthalmopathy, 2. Smoking patients with