Cushing's syndrome in pregnancy: a case report and mini review of the literature.

Adrenal diseases in pregnant women are diagnosed relatively rarely. The main cause of hypercortisolemia during pregnancy is Cushing's syndrome related to adrenal adenoma. It is important to diagnose Cushing's syndrome in pregnant women because it can lead to significant maternal and foetal complications and morbidity. However, due to physiological endocrine changes and symptoms in pregnant women the diagnosis of this disorder can be a challenge. One current case describes a 38-year-old pregnant woman with hypertension, oedema and an adrenal tumour. At the beginning, Conn syndrome was suspected, but after careful analysis Cushing's syndrome (with an adenoma of the right adrenal gland) was diagnosed. After delivery and 5 weeks of pharmacological treatment the patient underwent right side adrenalectomy by laparoscopy.

Increased serum osteoprotegerin in patients with primary adrenal insufficiency receiving conventional hydrocortisone substitution.

Patients treated for primary adrenal insufficiency (PAI) are at risk of steroid over-replacement, which may affect their skeleton. The study was aimed to investigate the effect of steroid substitution on serum osteoprotegerin and receptor activator of nuclear factor kappa-beta ligand (RANKL) levels in relation to bone mineral density (BMD) in PAI. Eighty patients (mean age 47.2±14.5 years, mean hydrocortisone dose 0.49±0.14 mg/kg/day) and 63 healthy subjects were included. Serum osteoprotegerin, RANKL, 25-hydroxyvitamin D3, calcium, phosphate, alkaline phosphatase, intact parathormone, and dehydroepiandrosterone-sulfate levels were evaluated in patients and controls. BMD was assessed in affected subjects using dual-energy X-ray absorptiometry. Mean osteoprotegerin concentration in PAI patients appeared significantly higher vs. controls (p=0.002), while RANKL levels were similar (p=0.430). Serum osteoprotegerin increased with age (p<0.001), but showed no correlation with daily hydrocortisone dose. Osteoprotegerin was negatively correlated with serum dehydroepiandrosterone-sulfate (p=0.008) and with BMD at the lumbar spine (p<0.001) and femoral neck (p=0.003). RANKL correlated negatively with PAI duration (p=0.029) and positively with daily hydrocortisone dose (p=0.018). Lumbar spine osteoporosis and osteopenia were found in 12 and 31 patients, respectively, whereas in femoral neck: in 5 and 33 individuals. Patients with osteoporosis displayed higher osteoprotegerin levels, but after the age-adjustment the correlation was lost. In conclusion, increased osteoprotegerin in PAI might reflect a compensatory response to enhanced bone resorption due to exogenous steroid excess and/or result from a deficit in adrenal androgens. RANKL levels remain within normal range on standard steroid replacement.

Thyroid hormones affect plasma ghrelin and obestatin levels.

Using radioimmunoassay, the effects of thyroid hormones on plasma total ghrelin (Gh) and obestatin (Ob) concentrations were evaluated in thyrotoxic patients with an excess of thyroid hormones and in hypothyroid patients lacking endogenous thyroid hormones. 24 patients with thyrotoxicosis, 25 hypothyroid patents after total thyreoidectomy performed due to thyroid cancer, and 17 control subjects were examined. Compared with the controls, the ghrelin and obestatin were elevated in hypothyroidism, while they were decreased in thyrotoxicosis. The plasma Gh and Ob levels differ depending on the thyroid function. In thyroid hormones deficiency, plasma Gh and Ob are increased, while in patients with excess of thyroid hormones, the levels of both Gh and Ob are definitely lower. Gh/Ob ratio is higher in hypothyroidism than in control subjects and thyrotoxic patients.

Association of the CYP27B1 C(-1260)A polymorphism with autoimmune Addison's disease.

Autoimmune Addison's disease (AAD) is a complex endocrine disorder with several susceptibility loci. This study was aimed to investigate the associations of CYP27B1 C(-1260)A and PDCD1 G7146A polymorphisms with AAD in a Polish cohort, comprising 101 AAD patients and 251 healthy controls. CYP27B1 encodes 1alpha-hydroxylase, responsible for conversion of the vitamin D (3) precursor into its active form, involved in the immune function. PDCD1 gene gives rise to an inhibitory immune receptor, expressed on activated lymphocytes. Polymorphic variants of these genes had previously been associated with various autoimmune disorders. Genotyping was performed by PCR-RFLP method. The CYP27B1 C(-1260) allele appeared significantly more frequent in AAD compared to controls ( P=0.020), yielding an OR of 1.53 (95% CI 1.07-2.19). The distribution of C(-1260)A genotypes also demonstrated significant difference ( P=0.003). Stratification according to the presence of concomitant autoimmune disorders revealed an association of the C(-1260) allele with the polyendocrine cases of AAD ( P=0.031), while no significance was found for the isolated ADD compared with healthy controls ( P=0.253). Overall, the association between AAD and C(-1260)A was confirmed in a meta-analysis of 325 AAD patients and 952 controls from three different European populations. Under a fixed-effect model, C(-1260) allele and CC genotype were associated with AAD susceptibility with a pooled OR of 1.44 (95% CI 1.18-1.75) and 1.88 (95% CI 1.42-2.36), respectively. No differences were observed for the PDCD1 G7146A between affected subjects and controls (p>0.05). In conclusion, this study confirms the association of the CYP27B1 C(-1260)A polymorphism with AAD, whereas the contribution of PDCD1 G7146A seems less likely.

The influence of growth hormone (GH) therapy on cardiac performance in patients with childhood onset GH deficiency.

OBJECTIVE: There is accumulating evidence that growth hormone (GH) plays an important role in the maintenance of normal cardiac growth and function. Abnormalities in left ventricular diastolic function and impairment of systolic function have also been reported in patients with GHD. In this study, we investigated the effects of 12 months GH replacement therapy on cardiac functional indices measured by echocardiography, the ECG stress test and SPECT imaging. DESIGN: Sixteen patients with childhood onset GHD (age 42.3+/-13.1 years, 10 males) were investigated before, and after, 12 months of GH treatment at a dosage of 0.02 IU/kg/day (7 microg/kg/day). The GH administration resulted in serum IGF-I levels within the normal range in all the patients. The following investigations were performed initially and after 12 months: electrocardiography, systolic and diastolic blood pressure, heart rate measurement, a complete Doppler-echocardiographic examination, treadmill exercise test and Technetium-99m sestamibi single-photon emission computer tomography (SPECT) imaging at rest and after exercise. RESULTS: Echocardiography showed improvement in left ventricular systolic function after GH treatment. End-systolic volume fell from 29.9+/-12.4 to 24.4+/-6.9 ml (p<0.05) and the ejection fraction increased from 56.2+/-7.2% to 63.2+/-6,1% (p<0.01). Left ventricular diameter and wall thickness did not change after GH treatment, although systolic increase in interventricular septum thickness (IVS%) and systolic increase in posterior wall thickness (PWT%) increased significantly (IVS% 52.2+/-31.9% vs. 67.3+/-30.4% and PWT% 48.7+/-20.2% vs. 58.0+/-17.7%, p<0.01, p<0.01, respectively). Contractile function, measured at midwall level, improved as left ventricular midwall fractional shortening (MWS) increased (16.11+/-6.55 vs. 23.30+/-5.89 %, p<0.01) and stress-corrected MWS increased between the examinations performed before and after 12 months of GH treatment (90.97+/-36.66 vs. 133.10+/-32.84 %, p<0.01). Diastolic function did not change, as assessed by early diastolic flow (E), diastolic flow secondary to atrial contraction (A), or the E/A ratio. The LV-mass index did not change significantly after GH treatment (78.4+/-22.1 vs. 81.9+/-21.1g/m(2)). After 12 months of GH treatment the myocardial performance index (MPI) decreased significantly from 0.483+/-0.146 at baseline to 0.410+/-0.086 at the end of the study (p<0.05). There was a trend towards an increase in exercise duration and capacity after GH treatment but the differences did not reach levels of statistical significance. SPECT imaging basally and after 12 months showed normal myocardial perfusion at rest and after exercise in all the patients. In conclusion, GH replacement therapy in adults with GHD demonstrated the beneficial effects on cardiac functions.

[Detection of pituitary autoantibody in patients with pituitary tumors and hypopituitarism]. / Wykrywanie autoprzeciwcial przysadkowych u chorych z gruczolakami przysadki i w niedoczynnosci przysadki.

UNLABELLED: In 80 patients, 73 cases of pituitary tumours and 7 cases of hypopituitarism, we performed pituitary autoantibodies assays in serum samples because in our previous studies we had found a high prevalence of pituitary autoantibodies in several autoimmune endocrine disorders. To detect the presence of pituitary autoantibodies we applied 2 methods, radioimmunoassay (RIA) and immunoblotting. The RIA was performed by solid phase technique in human pituitary microsome-coated polyethylene tubes. Following incubation with diluted sera of the patients labelled 125I-protein A was added to the tubes to detect the retained antibodies. In the sera of 33 patients we detected the presence of antibodies; in the other 47 patients no antibodies were found. The majority of the patients with positive antibody results were previously treated by pituitary irradiation. To evaluate the molecular weights of pituitary autoantigens the microsomal proteins were separated on SDS PAGE, then electrophoretically transferred to nitrocellulose membranes and reacted with diluted sera of 30 antibody-positive patients. The nitrocellulose strips were incubated with labelled 125I-protein A and autoradiographed. Using immunoblotting, 13 out of these 30 patients we found autoantibodies reacting with pituitary microsomal antigens of different molecular weights, most frequently reacting with a 68 kDa autoantigen. CONCLUSIONS: The prevalence of pituitary autoantibodies in patients with pituitary diseases is 41% lower than in autoimmune endocrine diseases. Pituitary autoantibodies usually appear in patients after pituitary irradiation or after neurosurgery followed by irradiation, but occur rarely in untreated patients with pituitary adenomas.

Expression of thyrotropin receptor (TSH-R), thyroglobulin, thyroperoxidase, and calcitonin messenger ribonucleic acids in thyroid carcinomas: evidence of TSH-R gene transcript in medullary histotype.

We studied the expression of the TSH receptor (TSH-R), thyroglobulin (Tg), thyroperoxidase (TPO), and calcitonin (CT) genes in a total of 53 tissues from 30 patients with thyroid carcinoma and from 9 patients with benign thyroid diseases. By Northern blot analysis of total RNA preparations, CT mRNA was expressed in all cases (n = 6) of medullary thyroid carcinoma (MTC). Surprisingly, 3 of them expressed the TSH-R mRNA, in association with the Tg and TPO mRNAs in 1. The presence of the TSH-R transcript in the neoplastic C-cells was confirmed in 1 MTC by in situ hybridization using a mixture of 3 oligonucleotide probes derived from dog TSH-R cDNA. With various degrees of expression, all differentiated thyroid carcinomas (20 papillary and 2 follicular) expressed TSH-R, Tg, and TPO, but not CT mRNAs. On the contrary, samples from 2 patients with anaplastic carcinoma did not express TSH-R, Tg, or TPO mRNA, but 1 of them expressed CT mRNA. All of the transcripts obtained from thyroid carcinomas (both primary and metastatic) were of the same size as the transcripts from normal or benign thyroid tissues, with the exception of 2 cases of differentiated thyroid cancer, in which TSH-R mRNA of lower mol wt (approximately 4.0 kilobases) was found in the absence of alteration in cDNA size and restriction map. The main conclusions of our study are that 1) the TSH-R gene is expressed in some MTC, which supports, at molecular level, the hypothesis of the existence of mixed follicular-medullary thyroid tumors; and 2) the expression of TSH-R, Tg, and TPO in undifferentiated thyroid cancer is lost.

Polyendocrine autoimmunity in thyroid diseases.

Anti-adrenal and anti-pituitary autoantibodies have been determined in 45 patients with autoimmune diseases of the thyroid, including 25 patients with Graves' disease and 20 patients with hypothyroidism of autoimmune origin. The determinations were carried out with the use of solid-phase RIA methods previously developed by us, involving polyethylene tubes coated with the solubilized microsomal fractions obtained from human adrenal and pituitary glands. In the majority of patients with autoimmune diseases of the thyroid the presence of both anti-adrenal and anti-pituitary autoantibodies was detected. In 13 among 20 patients with hypothyroidism of autoimmune origin, the presence of anti-adrenal autoantibodies, and in 12 the presence of anti-pituitary autoantibodies was found. Among 25 patients with Graves' disease, 19 had both anti-adrenal and anti-pituitary autoantibodies. In the majority of patients with autoimmune diseases of the thyroid the titers of autoantibodies of both types were high but in no case were the clinical symptoms of adrenal or pituitary hypofunction observed. Our studies indicate that the thyroid diseases of autoimmune origin can be regarded as manifestation of some more generalized autoimmunization process.

[Radioimmunologic determination of pituitary autoantibodies in patients with Addison's disease]. / Radioimmunologiczne oznaczanie autoprzeciwcial przysadkowych w chorobie Addisona.

The occurrence of pituitary autoantibodies was investigated in 34 patients, 25 females and 9 males, with Addison's disease of autoimmune origin and in 10 patients with tuberculous etiology of the disease. In both groups adrenal autoantibodies were also determined. Autoantibodies were determined by radioimmunoassay using solid phase technique in tubes coated with proteins of human adrenal and pituitary microsomal fractions. Pituitary autoantibodies were detected in 28 of 34 patients (20 females and 8 males) with autoimmune Addison's disease but only in 3 patients (2 females and one male) with nonimmune etiology of the disease. Both antipituitary and antiadrenal antibodies remain present in the circulation for many years as they were detected in autoimmune Addison's disease even after 10 or more years after the onset of the disease. There was a close relationship between the occurrence of pituitary antibodies and adrenal autoantibodies: they both appeared in the same patients and high titres of pituitary autoantibodies were associated with high titres of adrenal autoantibodies. This suggests a close similarity or identity of pituitary and adrenal autoantigens.

Measurement of TSAb directly in serum using FRTL-5 cells.

FRTL-5 cells were shown to be suitable for the measurement of thyroid stimulating antibody (TSAb) present in sera of patients with Graves' disease. Current methods for the assay of TSAb require the separation of immunoglobulin G (IgG) from patient sera. In this report the possibility to measure TSAb directly on serum was evaluated using FRTL-5 cells. To this purpose cells were seeded in 96-well plates and cultured for 4 days in medium deprived of TSH. Using this system bovine TSH was able to produce a significant stimulation of cAMP production at 1 microU/ml. Whole normal serum completely inhibited the stimulation of TSH as well as that of TSAb, while diluted serum was devoid of any effect. Heat inactivated sera and IgGs, prepared by DEAE Sephadex separation, were diluted in hypotonic medium and incubated with cells for 1 h at 37 C. After incubation cAMP was measured in the assay medium by RIA. In some experiments the effects of graded dilutions of sera and IgGs with known TSAb activity were compared. Sera as well as IgGs increased the cAMP production, but, at the highest concentrations, an inhibitory effect was evident. For this reason sera were tested after appropriate dilution. Thirteen/27 (48%) sera and 22/27 (81%) IgGs from patients with Graves' disease were TSAb positive. The effect of Graves' sera on adenylate cyclase stimulation was completely inhibited by an anti-human IgG. The results of stimulation produced by Graves' sera and IgGs were highly correlated (r = 0.97, p less than 0.001). In conclusion it is possible to measure TSAb directly in serum using FRTL-5 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

A radioimmunoassay for the detection of adrenal autoantibodies.

A solid phase radioimmunoassay for adrenal antibodies is described. In the assay plastic tubes coated with adrenal microsomes (100 micrograms/ml) were incubated with human sera diluted from 1:50 to 1:5000 and the retained antibodies detected by subsequent incubation with 125I-labelled protein A. The method was precise over the range of serum dilution of 1:250 to 1:5000. In the group of 30 patients with Addison's disease 19 had positive results in adrenal antibody radioimmunoassay (RIA). Comparative studies of RIA and immunofluorescence (IFL) revealed that there was partial correlation of adrenal antibody results in patients with high titre antibodies whereas RIA usually was more sensitive than IFL in patients with low titre antibodies. Computerized tomography (CT) displayed bilateral adrenal atrophy in most patients who had adrenal antibodies. On the other hand, patients with low RIA results and negative IFL antibodies had predominantly adrenal calcifications on scans.

Elevation of maternal serum myoglobin concentrations during delivery.

Serum myoglobin was determined by radioimmunoassay in 32 women immediately after delivery, in the umbilical cord blood of 30 newborn infants, in 18 women in the third trimester of pregnancy and in 24 healthy non-pregnant women. In maternal blood following delivery there was a 3 to 10 fold rise in serum myoglobin concentration (mean +/- SD = 117.2 +/- 82.5 ng/ml). On the other hand, in umbilical cord blood and in late pregnancy, serum myoglobin concentrations did not differ from those in healthy non-pregnant women (mean +/- SD = 23.6 ng/ml).