Renin, Aldosterone, and Cortisol in Pregnancy-Induced Hypertension.

INTRODUCTION: We aimed to assess renin, aldosterone, and cortisol in the early stages of pregnancy-induced hypertension (PIH), i. e., at the time of diagnosis. METHODS: During the postural test, we measured aldosterone, renin [Liason DiaSorin Inc. (Italy)], as well as cortisol, sodium, potassium, and 24-h urinary sodium and potassium excretion in 62 women with newly diagnosed PIH, 70 healthy women during the 3rd trimester of pregnancy, and in 22 healthy non-pregnant women. RESULTS: In all groups, there was a significant increase in aldosterone and renin in upright versus supine posture (p<0.01). Both supine and upright aldosterone concentrations were higher in healthy pregnant women than in women with PIH and the lowest in healthy not-pregnant [supine (median±intequartile range): 25.04±18.4 ng/dL, 18.03±12.58 ng/dL, and 7.48±4.78 ng/dL, p<0.001, upright: 31.60±21.32 ng/dL, 25.11±13.15 ng/dL, and 12.4±12.4 ng/dL, p<0.001, for healthy pregnant, pregnant with PIH, and non-pregnant, respectively]. Supine renin concentrations were higher only in healthy pregnant (p<0.001), while in the upright position, there was a difference only between healthy pregnant and women with PIH (p=0.002). Both in supine and upright positions, there was no difference in the aldosterone-to-renin ratio between healthy pregnant women and women with PIH, though, in both groups, the ratio was higher than in non-pregnant women (p<0.001). Morning cortisol concentrations and 24-h urinary sodium excretion were lower in women with PIH than in healthy pregnant (p<0.001, p=0.002, respectively). CONCLUSION: Hyperaldosteronism is not involved in the etiology of PIH. In PIH, there is also a tendency towards lower sodium excretion and lower morning cortisol concentrations.

Limited Role of Endogenous Vasopressin/Copeptin in Stimulation of ACTH-Cortisol Secretion during Glucagon Stimulation Test in Humans.

Copeptin is a stable part of a vasopressin precursor that closely mirrors arginine vasopressin (AVP) secretion. It is known that AVP/copeptin is also released in response to nonosmotic stimuli, such as stress evoked during anterior pituitary dynamic testing. In order to examine the role of AVP in challenging the hypothalamo-pituitary-adrenal axis, we assessed adrenocorticotropic hormone (ACTH), cortisol, copeptin and growth hormone (GH) during a glucagon stimulation test (GST) in 10 patients with satisfactory initial cortisol concentrations (mean ± SD: 20.34 ± 5.10 µg/dL) and failure to show any further cortisol increment on stimulation. For comparison, we measured copeptin in two subjects during an insulin tolerance test (ITT). During GST, there was an increase in copeptin (p = 0.02, average individual increase of 98%, range 10% to 321%). There was a robust increase in GH (p = 0.002, average increase 3300%), a decline in cortisol (p = 0.02, average decline 21.8%) and a fall in ACTH (p = 0.06). The relative increase in copeptin during ITT (176% and 52.2%) overlapped with increments observed during GST; however, here there was an increase in cortisol (20.45→24.26 µg/dL and 4.23→29.29 µg/dL, respectively). There was a moderate correlation between copeptin and GH concentrations (r = 0.4235, p = 0.0007). These results confirm that AVP is not crucial for ACTH-cortisol stimulation, though it might be an important factor in GH secretion.

The effects of season (spring versus autumn) on diagnosis of normocalcemic primary hyperparathyroidism.

Background: Raised parathormone (PTH) and normal calcium concentrations can be observed both in normocalcemic primary hyperparathyroidism (nPHPT) and in secondary hyperparathyroidism, e.g. due to vitamin D deficiency. We assessed the impact of season on the validity of diagnosis of nPHPT in terms of screening investigations to be performed in the primary care settings. Material and methods: On two occasions (March/April & September/October) we measured vitamin D (25OHD), PTH and total calcium in 125 healthy subjects, age range 6-50, not taking any vitamin D supplements. Results: In autumn there was an increase in 25OHD concentrations (from 18.1 ± 7.37ng/ml to 24.58 ± 7.72ng/ml, p<0.0001), a decline in PTH from 44.40 ± 17.76pg/ml to 36.63 ± 14.84pg/ml, p<0.001), without change in calcium levels. Only 45 subjects (36%) were vitamin D sufficient (25OHD>20/ml) in spring versus 83 (66.4%) in autumn, p<0.001. Elevated PTH concentrations were noted in 10 subjects in spring (8%) and in six subjects (4.8%) (p<0.05) in autumn. In spring, however, eight out of ten of these subjects (80%) had 25OHD<20 ng/ml, versus one in six (16.7%) in autumn (p<0.01). Normalization of PTH was observed in seven out ten subjects (70%), and all of them had 25-OHD<20 ng/ml in spring. Conclusions: In spring elevated PTH concentrations in the setting of normocalcemia are more likely to be caused by 25OHD deficiency rather by nPHPT. In contrast, in autumn, increased PTH concentrations are more likely to reflect nPHPT. We postulate that screening for nPHPT should be done in 25OHD replete subjects, i.e. in autumn rather than in spring.

Establishing Reference Ranges for Aldosterone, Renin and Aldosterone-to-Renin Ratio for Women in the Third-Trimester of Pregnancy.

OBJECTIVE: Diagnosis of primary hyperaldosteronism in pregnancy is complicated due to lack of reference ranges for aldosterone, renin and aldosterone-to-renin ratio. We have endeavoured to establish third-trimester reference ranges for the above-mentioned parameters. DESIGN & PATIENTS: We performed postural tests for aldosterone and renin (chemiluminescence immunoassay Liason® DiaSorin Inc., Italy) in 70 healthy pregnant women (age 30.53±4.51 years), at 32.38±4.25 weeks of gestation and in 22 non-pregnant healthy women (age 33.08±8.72 years). RESULTS: Aldosterone reference ranges were 6.51-73.97 ng/dl and 12.33-86.38 ng/dl, for supine and upright positions, respectively and that for renin were 6.25-59.36 µIU/ml and 11.12-82.55 µIU/ml, respectively. Aldosterone and renin concentrations were higher in an upright position (p=0.000459 and p=0.00011, respectively). In contrast, aldosterone-to-renin ratio was not affected by posture (i. e. 0.497-3.084 ng/dl/µIU/ml versus 0.457-3.06 ng/dl/µIU/ml, p=0.12), but was higher (p=0.00081) than in non-pregnant controls. In comparison to manufacturer-provided non-pregnant reference range, supine aldosterone concentrations increased by 556% (lower cut-off) and 313% (upper cut-off), while upright aldosterone concentrations increased by 558% (lower cut-off) and 244% (upper cut-off). The reference range for supine renin concentrations increased by 223% (lower cut-off) and 48.7% (upper cut-off), while upright renin concentrations increased by 253% (lower cut-off) and 79% (upper cut-off). CONCLUSIONS: There is an upward shift in aldosterone and renin reference ranges in the third-trimester of pregnancy accompanied by an increase in an aldosterone-to-renin ratio, that is not influenced by posture. It remains to be established whether the aldosterone-to-renin ratio may be used as a screening tool for primary hyperaldosteronism in pregnancy.

Correction: C-Peptide and leptin system in dichorionic, small and appropriate for gestational age twins-possible link to metabolic programming?

The original version of this Article was updated shortly after publication to correct two mistakes.Under Figure 2, "The results were recalculated accordingly: nanograms [ng] of studied protein per 100 µ" should read "The results were recalculated accordingly: picograms [pg] of studied protein per 100 µg".

C-Peptide and leptin system in dichorionic, small and appropriate for gestational age twins-possible link to metabolic programming?

Children born small for gestational age (SGA) are at increased risk of future glucose intolerance and type 2 diabetes, possibly after due intrauterine metabolic programming. Soluble leptin receptor (SLR) limits leptin access to signal-transducing membrane receptors. The present study examines whether SGA and appropriate for gestational age (AGA) twins differ with regard to their C-peptide, glucose and leptin systems. The markers C-peptide, glucose, fetal leptin, and SLR in cord blood were assessed in children from dichorionic twin pregnancies at delivery. In 32 cases, weight differed by >15% between twins: one demonstrated Intrauterine Growth Retardation (IUGR) (<10th percentile-SGA), while the other did not (AGAI). The other 67 pairs presented appropriate weight for gestational age (AGAII). Placental leptin and placental leptin receptor content were also assessed. Despite the same concentrations of glucose, the SGA twins maintained a higher level of C-peptide [44.48 pmol/l vs. 20.91 pmol/l, p < 0.05] than the AGAI co-twins, higher HOMA index, calculated as [C-peptide] x [Glucose] (p = 0.045), in cord blood, and a higher level of SLR [SGA vs AGAI-mean: 28.63 ng/ml vs. 19.91 ng/ml, p < 0.01], without any differences in total leptin (p = 0.37). However, SGA placentas demonstrated higher leptin level [130.1 pg/100 g total protein vs 83.8 pg/100 g total protein, p = 0.03], without differences in placental leptin receptor (p = 0.66). SGA/IUGR twins demonstrate relative insulin resistance accompanied by decreased fetal and increased placental leptin signaling. We speculate that relative insulin resistance and changes in the leptin system might be the first evidence of processes promoting deleterious metabolic programming for post-natal life.

Subclinical thyroid dysfunction in the first trimester of pregnancy: 'Disease' versus physiological (pulsatile) variation in TSH concentrations.

BACKGROUND: There is no universal consensus regarding cut-off points for TSH in pregnancy, so concentrations of 2.5 or 4.0 mIU/L were suggested for first trimester (Endocrine Society [2012] and ATA [2017] guidelines, respectively). Yet, the impact of physiological variation in TSH secretion has not been assessed. SUBJECTS AND METHODS: We assessed baseline concentrations of free T4, free T3 and TSH at 30-minute intervals (between 7.00 and 9.00 hours) in 110 healthy pregnant women, age 30.2 ± 6.0 years, 9.9 ± 2.4 weeks of gestation, and in 19 female controls, age 28.9 ± 10.7. RESULTS: Mean TSH concentrations in pregnant women were 1.62 ± 1.26 mIU/L and on average varied by 39.5% (dispersion between the highest and the lowest TSH), with no difference in TSH variation between pregnant women and controls. Taking into account the highest TSH out of five consecutive measurements, TSH >2.5 mIU/L and TSH above 4.0 mIU/L were found in 23 (20.9%) and 10 (9.1%) pregnant women, respectively. In contrast, when the lowest TSH value was considered, then concentrations of TSH >2.5 mIU/L and >4.0 mIU/L were found in 14 (12.7%) and 4 (3.6%) women, respectively. This discrepancy was even more pronounced in aTPO-negative subjects (21 [21.2%] vs 8 [8.1%] women, for TSH >2.5 mIU/L, and six [6.06%] vs one [1.01%], for TSH >4.0 mIU/L). Furthermore, either six (5.4%) or 10 (9.1%) women had TSH concentrations below 0.1 mIU/L. CONCLUSIONS: In a significant number of patients, diagnosis of subclinical thyroid dysfunction could be erroneously made not as a result of 'disease', but as a result of physiological variation in TSH concentrations.

Adequate timing and constant supervision are the keys for successful implementation of levothyroxine or levothyroxine/paracetamol absorption test.

BACKGROUND: Levothyroxine (LT4) pseudomalabsorption due to medication non-adherence results in significant costs for Health Service. High dose LT4 or LT4/paracetamol absorption test is used in such cases. Hence, establishment of an optimal test protocol and timing of sample collection is of utmost importance. CASE PRESENTATION: A 34-year old woman was admitted to our Department because of severe hypothyroidism [on admission thyrotropin (TSH) > 100 µIU/ml, free thyroxine (FT4) 0.13 ng/dl (ref. range 0.93-1.7)] despite apparently taking 1000 µg of LT4 a day. Autoimmune hypothyroidism had been diagnosed 4 years before during post-partum thyroiditis. Subsequently, it was not possible to control her hypothyroidism despite several admissions to two University Hospitals and despite vehement denial of compliance problems. There was no evidence of coeliac disease or other malabsorption problems, though gluten-free and lactose-free diet was empirically instigated without success. A combined paracetamol (1000 mg)/LT4 (1000 µg) absorption test was performed in one of these Hospitals. This showed good paracetamol absorption (from < 2 µg/ml to 14.11 µg/ml at 120 min), with inadequate LT4 absorption (FT4 increase from 5.95 pmol/l to 9.92 pmol/l at 0 and 120 min respectively). About 2 years prior to admission to our Department the patient was treated with escalating doses of levothyroxine [up to 3000 µg of T4 and 40 µg of triiodothyronine (T3) daily] without significant impact on TSH (still > 75 µIU/ml, and FT4 still below reference range).After admission to our Department we performed a 2500 µg LT4 absorption test with controlled ingestion of crushed tablets, strict patient monitoring and sampling at 30 min intervals. We observed a quick and striking increase in FT4 from 0.13 to 0.46, 1.78, 3.05 and 3.81 ng/dl, at 0, 30, 60, 90 and 120 min, respectively. Her TSH concentration decreased to 13.77 µIU/ml within 4 days. When informed, that we had managed to "overcome" her absorption problems, she discharged herself against medical advice and declined psychiatric consultation. CONCLUSIONS: Adequate patient supervision and frequent sampling (e.g. every 30 min for 210 min) is the key for successful implementation of LT4 absorption test. Paracetamol coadministration appears superfluous in such cases.

Metformin paradoxically worsens insulin resistance in SHORT syndrome.

BACKGROUND: SHORT syndrome is an autosomal dominant condition associated severe insulin resistance (IR) and lipoatrophy due to post-receptor defect in insulin signaling involving phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), where no clear treatment guidelines are available. METHODS: We attempted to test the efficacy metformin in a female patient with SHORT syndrome by measuring glucose and insulin during an extended Oral Glucose Tolerance Test (OGTT) in a 21-year old patient (BMI 17.5 kg/m2), who presented for endocrine assessment with a history of amenorrhoea. RESULTS: She had lipid concentrations within the reference range, normal thyroid function tests, prolactin, gonadotropins, estradiol and androgens with Free Androgen Index 4.52. Extended Oral Glucose Tolerance Test was performed and showed severe IR. She was then started on metformin 850 mg twice a day, and had repeated OGTT. This showed dramatic worsening of glucose tolerance (e.g. glucose 96 mg/dl versus 187 mg/dl and 68 mg/dl versus 204 mg/dl at 120 and 150 min of OGTT, respectively). This was accompanied by a massive increase of already high insulin concentrations (e.g. from 488.6 to > 1000 µIU/ml, and from 246.8 to > 1000 µIU/ml at 120 and 150 min of OGTT, respectively). Insulin concentrations remained above upper assay detection limit also at 180 min of OGTT on metformin treatment (> 1000 µIU/ml versus 100.6 µIU/ml without metformin). CONCLUSIONS: Metformin treatment may paradoxically lead to deterioration of insulin resistance and to development of glucose intolerance in SHORT syndrome. Hence, metformin treatment might be potentially harmful in these patients. Though, the precise cause of such profound and paradoxical worsening of glucose tolerance post metformin remains unknown, SHORT syndrome might prove to be an interesting model to study the mechanism(s) of metformin action.

How much insulin resistance in polycystic ovary syndrome? Comparison of HOMA-IR and insulin resistance (Belfiore) index models.

INTRODUCTION: Polycystic ovary syndrome (PCOS), the commonest endocrinopathy of women in reproductive age, is often accompanied by insulin resistance (IR), hirsutism and/or fertility problems. The aim of the study was to assess the prevalence of IR in women diagnosed with PCOS. MATERIAL AND METHODS: The study involved 137 women diagnosed with PCOS, according to the Rotterdam consensus criteria (2003). Insulin resistance was assessed according to the HOMA-IR method and insulin resistance (Belfiore) index (IRI) derived from glucose and insulin during the oral glucose tolerance test. RESULTS: There was a significant (p < 0.0001) but relatively moderate correlation between IRI and HOMA-IR (r = 0.5 and r = 0.57 for a linear and non-linear model, respectively). Insulin resistance was more prevalent according to IRI (49.6%) than according to HOMA-IR (22.6% and 15.8% for 3.46 and 3.8 cut-off points, respectively, p < 0.01). The majority of patients with high HOMA-IR also had high IRI (e.g. 86%, for HOMA > 3.8), but the majority of patients with raised IRI would not be diagnosed as insulin resistant according to HOMA (61.7% and 73.5%, for HOMA-IR3.46 and HOMA-IR3.80, respectively). CONCLUSIONS: The insulin resistance (Belfiore) index indicates more cases of insulin resistance than HOMA-IR in women with PCOS. Therefore, detection of insulin resistance among women with PCOS is highly method-dependent with more severe cases being detected with HOMA-IR than with IRI.

Prevalence of Dyslipidaemia and Pre-Diabetes Among Women with Polycystic Ovary Syndrome (PCOS): Do We Overestimate Cardiovascular Risk?

PCOS is widely accepted as associated with an increased cardiovascular risk, however, without convincing evidence of an increased cardiovascular mortality. We assessed prevalence of obesity, glucose intolerance, and dyslipidaemia in 490 women with PCOS, aged 24.75±8.05 years, diagnosed according to the Rotterdam consensus criteria. Fifty-two percent of women had BMI<26 kg/m2, 81.8% had total cholesterol<200 mg/dl, 82.8% had LDL cholesterol<130 mg/dl (48.3%<100 mg/dl), 81.4% had triglycerides<150 mg/dl, 96.08% had fasting glucose<100 mg/dl, 90.3% had glucose<140 mg/dl at 120' of OGTT. The most frequent abnormality was low HDL cholesterol, as only 33.9% had LDL>60 mg/dl. Combination of several risk factors related to dyslipidaemia was, however, relatively rare, for example, a combination of raised total cholesterol and LDL cholesterol was present only in 2.9% of subjects. An increase in BMI, total cholesterol, LDL-cholesterol, and glucose concentrations at 120' of OGTT was more pronounced in women, who had raised concentrations of at least two androgens (n=172, 35.1%), yet there was no increase in insulin resistance parameters, that is, HOMA-IR, QUICKI, McAuley, or Belfiore index. Contrary to common belief, over 50% of women with PCOS have normal body weight, and with exception of lower HDL cholesterol, most have no significant dyslipidaemia or glucose intolerance. Women with normal or borderline abnormal androgens, who form the majority of PCOS subjects, seem to have more healthy metabolic profile. This might be one of the reasons for the absence of evidence of an increased CV mortality in women with PCOS.

Limitations of insulin resistance assessment in polycystic ovary syndrome.

BACKGROUND: Though insulin resistance (IR) is common in polycystic ovary syndrome (PCOS), there is no agreement as to what surrogate method of assessment of IR is most reliable. SUBJECTS AND METHODS: In 478 women with PCOS, we compared methods based on fasting insulin and either fasting glucose (HOMA-IR and QUICKI) or triglycerides (McAuley Index) with IR indices derived from glucose and insulin during OGTT (Belfiore, Matsuda and Stumvoll indices). RESULTS: There was a strong correlation between IR indices derived from fasting values HOMA-IR/QUICKI, r = -0.999, HOMA-IR/McAuley index, r = -0.849 and between all OGTT-derived IR indices (e.g. r = -0.876, for IRI/Matsuda, r = -0.808, for IRI/Stumvoll, and r = 0.947, for Matsuda/Stumvoll index, P < 0.001 for all), contrasting with a significant (P < 0.001), but highly variable correlation between IR indices derived from fasting vs OGTT-derived variables, ranging from r = -0.881 (HOMA-IR/Matsuda), through r = 0.58, or r = -0.58 (IRI/HOMA-IR, IRI/QUICKI, respectively) to r = 0.41 (QUICKI/Stumvoll), and r = 0.386 for QUICKI/Matsuda indices. Detailed comparison between HOMA-IR and IRI revealed that concordance between HOMA and IRI was poor for HOMA-IR/IRI values above 75th and 90th percentile. For instance, only 53% (70/132) women with HOMA-IR >75th percentile had IRI value also above 75th percentile. There was a significant, but weak correlation of all IR indices with testosterone concentrations. CONCLUSIONS: Significant number of women with PCOS can be classified as being either insulin sensitive or insulin resistant depending on the method applied, as correlation between various IR indices is highly variable. Clinical application of surrogate indices for assessment of IR in PCOS must be therefore viewed with an extreme caution.

Addison's disease concomitant with corticotropin deficiency and pituitary CRH resistance - a case report.

A 36-year-old woman was found to have a low morning ACTH concentration despite a history of Addison's disease. Past medical history: At the age of 23 years the subject developed Graves's disease, which was treated with radioiodine. At about the same time, she claimed to have two episodes of pancreatitis treated with cholecystectomy. About seven months later she was euthyroid on L-thyroxine (TSH 1.51 mIU/mL) but was admitted with hypotension, hyponatraemia (sodium 109 mmol/L), and low morning cortisol (119 nmol/L). Further investigations confirmed primary adrenal failure with ACTH concentration of 779 pg/mL (ref. range 0-60) prior to the dose of hydrocortisone. About nine years later she complained about tiredness. Clinically she was normotensive and not pigmented. BMI 22.3 kg/m². Periods were regular. ACTH concentration was surprisingly low (ACTH 8.53 pg/mL, ref. range 0-46), despite very low cortisol (3.37 nmol/L). She was admitted for further assessment. INVESTIGATIONS: Pituitary MRI scan was unremarkable. An insulin tolerance test was performed and showed a clear increase of ACTH (from 15.2 to 165 pg/mL). There was, however, hardly any increase of ACTH after CRH stimulation (from 6.05 pg/mL to 10.2 pg/mL), thus demonstrating central CRH resistance. In summary, this patient developed secondary adrenal failure in the setting of previous Addison's disease. Interestingly, hypoglycaemia (but not CRH) provided a stimulus for ACTH release, thus demonstrating CRH resistance. The case confirms that besides CRH, other factors are responsible for stimulation of the ACTH-cortisol axis during insulin tolerance test.

Copeptin as a marker of an altered CRH axis in pituitary disease.

BACKGROUND: Copeptin (pre-proAVP) secreted in equimolar amounts with vasopressin closely reflects vasopressin release. Copeptin has been shown to subtly mirror stress potentially mediated via corticotrophin-releasing hormone. To further test a potential direct interaction of corticotrophin-releasing hormone with copeptin release, which could augment vasopressin effects on pituitary function, we investigated copeptin response to corticotrophin-releasing hormone. PATIENTS AND METHODS: Cortisol, adrenocorticotropin and copeptin were measured in 18 healthy controls and 29 subjects with a history of pituitary disease during standard corticotrophin-releasing hormone test. RESULTS: Patients with previous pituitary disease were subdivided in a group passing the test (P1, n = 20) and failing (P2, n = 9). The overall copeptin response was higher in controls than in subjects with pituitary disease (area under the curve, p = 0.04 for P1 + P2) with a maximum increase in controls from 3.84 ± 2.86 to 12.65 ± 24.87 pmol/L at 30 min, p < 0.05. In contrast, both groups of pituitary patients lacked a significant copeptin response to corticotrophin-releasing hormone, and even in P1, where adrenocorticotropin concentrations increased fourfold (mean, 21.48 vs. 91.53 pg/mL, p < 0.01), copeptin did not respond (e.g., 4.35 ± 5.81 vs. 5.36 ± 6.79 pmol/L, at 30 min, p = ns). CONCLUSIONS: Corticotrophin-releasing hormone is able to stimulate copeptin release in healthy controls suggesting a direct interaction of corticotrophin-releasing hormone and vasopressin/vasopressin. Interestingly, this relation is altered already in the group of pituitary patients who pass the standard corticotrophin-releasing hormone test indicating (1) the corticotrophin-releasing hormone-adrenocorticotropin-cortisol response is largely independent from the vasopressin system, but (2) the corticotrophin-releasing hormone-vasopressin interaction reflected by copeptin may be much more sensitive to reveal subtle alterations in the regulation of pituitary function.

Copeptin under glucagon stimulation.

Stimulation of growth hormone (GH) and adrenocorticotropic hormone (ACTH) secretion by glucagon is a standard procedure to assess pituitary dysfunction but the pathomechanism of glucagon action remains unclear. As arginine vasopressin (AVP) may act on the release of both, GH and ACTH, we tested here the role of AVP in GST by measuring a stable precursor fragment, copeptin, which is stoichiometrically secreted with AVP in a 1:1 ratio. ACTH, cortisol, GH, and copeptin were measured at 0, 60, 90, 120, 150, and 180 min during GST in 79 subjects: healthy controls (Group 1, n = 32), subjects with pituitary disease, but with adequate cortisol and GH responses during GST (Group 2, n = 29), and those with overt hypopituitarism (Group 3, n = 18). Copeptin concentrations significantly increased over baseline 150 and 180 min following glucagon stimulation in controls and patients with intact pituitary function but not in hypopituitarism. Copeptin concentrations were stimulated over time and the maximal increment correlated with ACTH, while correlations between copeptin and GH were weaker. Interestingly, copeptin as well as GH secretion was significantly attenuated when comparing subjects within the highest to those in the lowest BMI quartile (p < 0.05). Copeptin is significantly released following glucagon stimulation. As this release is BMI-dependent, the time-dependent relation between copeptin and GH may be obscured, whereas the close relation to ACTH suggests that AVP/copeptin release might be linked to the activation of the adrenal axis.

Familial partial lipodystrophy as differential diagnosis of polycystic ovary syndrome.

UNLABELLED: According to current diagnostic criteria, polycystic ovary syndrome (PCOS) is effective as a diagnosis of exclusion. Here, we present a case of a 31-year-old woman with a history of oligomenorrhoea and hirsutism, who, despite a "muscular" appearance and a normal body mass index (22.27 kg/m2), was found to have an extreme insulin resistance and diabetes accompanied by hyperandrogenism and polycystic ovaries. An autoimmune screen for possible latent autoimmune diabetes in adults was negative. She was subsequently found to have familial partial lipodystrophy (FPLD2, OMIM #151660) caused by an R482Q mutation in the LMNA gene encoding lamin A/C. This mutation results in arginine to glutamine substitution at the protein level, while phenotypically this condition presents with a loss of body fat, insulin resistance, dyslipidaemia, and other features mimicking PCOS. Interestingly her mother, with a history of myocardial infarction and diabetes at the age of 46 but no oligomenorrhoea, was also found to harbour the same mutation (LMNA R482Q). CONCLUSIONS: Our case highlights the importance of assessment of adipose tissue distribution, as well as a significance of assessment of glucose tolerance and insulin resistance in the differential diagnosis of PCOS. Furthermore, patients with atypical adipose tissue distribution should be referred for formal genetic testing.

A single bolus of high dose levothyroxine (L-T4) as a test in cases of suspected poor compliance to L-T4 therapy.

BACKGROUND: Though compliance (or adherence) problems, as well as inappropriate levothyroxine (L-T4) intake (e.g. with meal, other drugs or certain foods that can significantly affect absorption) are very common, the issue is often either not mentioned or even frankly denied by patients. CASE PRESENTATION: We describe three cases of patients who presented with high TSH (ranging from about 30 to 200 mIU/l), with concomitantly either high, normal or low free thyroxine (FT4), despite treatment with high doses of L-T4. The above mentioned problems with adjustment of L-T4 dose persisted for several months or even years. Coeliac disease screen was negative in all cases. In all these patients administration of a single bolus of L-T4 (1000 µg) or two doses of 1000 µg of L-T4 within 48 h resulted in a quick increase in FT4 (thus confirming proper absorption) and in normalization of TSH within a week. No adverse effects of administration of these high doses of L-T4 were observed. CONCLUSIONS: Our data support the efficacy, as well as safety of administration of single bolus of high dose L-T4 as a test for possible compliance/adherence problems.

Mechanisms of Normalisation of Bone Metabolism during Recovery from Hyperthyroidism: Potential Role for Sclerostin and Parathyroid Hormone.

Sclerostin, a protein expressed by osteocytes, is a negative regulator of bone formation. The aim of the study was to investigate the relationship between parathyroid hormone (PTH) and markers of bone metabolism and changes of sclerostin concentrations before and after treatment of hyperthyroidism. Patients and Methods. The study involved 33 patients (26 women), age (mean ± SD) 48 ± 15 years, with hyperthyroidism. Serum sclerostin, PTH, calcium, and bone markers [osteocalcin (OC) and collagen type I cross-linked C-telopeptide I (CTX)] were measured at diagnosis of hyperthyroidism and after treatment with thiamazole. Results. After treatment of hyperthyroidism a significant decrease in free T3 (FT3) and free T4 (FT4) concentrations was accompanied by marked decrease of serum sclerostin (from 43.7 ± 29.3 to 28.1 ± 18.4 pmol/L; p < 0.001), OC (from 35.6 ± 22.0 to 27.0 ± 14.3 ng/mL; p < 0.001), and CTX (from 0.49 ± 0.35 to 0.35 ± 0.23 ng/dL; p < 0.005), accompanied by an increase of PTH (from 29.3 ± 14.9 to 39.8 ± 19.8; p < 0.001). During hyperthyroidism there was a positive correlation between sclerostin and CTX (r s = 0.41, p < 0.05) and between OC and thyroid hormones (with FT3 r s = 0.42, with FT4 r s = 0.45, p < 0.05). Conclusions. Successful treatment of hyperthyroidism results in a significant decrease in serum sclerostin and bone markers concentrations, accompanied by an increase of PTH.

Spuriously high androstendione concentrations due to assay interference as a cause of diagnostic conundrum in women with oligomenorrhoea.

UNLABELLED: Polycystic ovary syndrome (PCOS) is a diagnosis of exclusion. We present two cases of women with oligomenorrhoea and high concentration of androstendione, suggestive of possible androgen-secreting tumour; caused by assay interference. The first patient, investigated for oligomenorrhoea, had no significant hirsutism or acne. Androstendione concentration was above 10.0 ng/ml (rr: 0.3-3.3 ng/ml). In order to rule out possible androgen-secreting tumour or hypercortisolaemia we performed 48-hour low dose dexamethasone suppression test (LDDST). This failed to demonstrate adequate suppression of androstendione (6.05 ng/ml and 9.32 ng/ml after the first and the second day respectively). Pelvic ultrasound examination showed polycystic ovaries, while abdominal CTscan failed to show any ovarian or adrenal lesion. Despite such high androstendione concentrations, urinary steroid profile (gas chromatography/mass spectrometry method) yielded normal results. Hence a possibility of androstendione assay interference was raised. The second patient was also admitted for investigations of oligomenorrhoea. Clinical examination was unremarkable. There was a high concentration of testosterone 0.78 ng/ml (rr. 0.084-0.481 ng/ml) and androstendione above 10.0 ng/ml (rr: 0.3-3.3 ng/ml). LDDST failed to demonstrate any suppression of androstendione, while recalculated concentrations of androstendione after serial dilutions were markedly lower in comparison to initial values. Therefore, such high androstendione concentrations (i.e. above the upper limit of the assay) must have resulted from assay interference. In both cases a final diagnosis of PCOS was established. CONCLUSIONS: In the absence of clinical features, contrasting with unusually high androgen levels, a possibility of androgen assay interference should be considered in differential diagnosis of hyperandrogenism or PCOS.

Effects of maternal vitamin D status on pregnancy outcomes, health of pregnant women and their offspring.

OBJECTIVE: Pregnancy increases the demand for vitamins, including vitamin D. Data on effects of vitamin D deficiency for pregnant woman and fetus available in Poland are scarce. The aim of this study was to evaluate vitamin D3 concentration in pregnant women and its influence on pregnancy course, health of pregnant women and their offspring. PATIENTS AND METHODS: The study included 102 healthy pregnant women, aged 21 to 40 years, mean 30.5±4.9 years. Women were divided into three groups based on 25(OH)D serum concentration in the third trimester of pregnancy: Group I - with sufficient 25(OH)D serum concentration (>30 ng/ml), Group II - with vitamin D3 insufficiency (20-30 ng/ml), Group III - with serious vitamin D deficiency (<20 ng/ml). RESULTS: Optimal vitamin D concentrations were found only in 31.2% of women, however in winter months only in 16%. Bacterial vaginosis was significantly more common in women with vitamin D deficiency and insufficiency (p<0.05). In contrast, there were no relations between vitamin D status and the incidence of gestational diabetes, preeclampsia, mode of delivery and size of newborns. A relationship between vitamin D deficiency and insufficiency during pregnancy and subsequent incidence of respiratory infections in children (p<0.05) was demonstrated. CONCLUSIONS: 1. The current model of vitamin D supplementation in pregnant women in Poland is insufficient, particularly in winter. 2. Vitamin D deficiency in pregnant women fosters development of bacterial vaginosis during pregnancy and recurrent respiratory infections in children, suggestive of the role of vitamin D in prevention of infections.