bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-ribosomal antibodies from lupus patients bind DNA
Authors: CAPONI L , CHIMENTI D , PRATESI F , MIGLIORINI P
Published: March 2003

Antibodies specific for ribosomal P proteins (anti-P) are a hallmark of systemic lupus as anti-DNA antibodies are. It has been reported that anti-P antibodies are more frequently detected in anti-dsDNA positive sera. The aim of the present study was to verify the binding ability of anti-P antibodies towards polynucleotides and nucleosomes. We purified anti-P antibodies from 8 SLE patients' sera and we analysed them by ELISA on plates coated with DNA or nucleosomes. We performed also inhibition experiments in order to verify the specificity of the binding. All the purified anti-P antibodies bound DNA, but some anti-DNA binding activity remained among the non-anti-P antibodies in the flow through. Only half of the purified antibodies bound to nucleosomes, and anti-nucleosomal activity was demonstrated also in non anti-P antibody fraction. The inhibition experiments performed on two anti-P antibodies pointed out that only the homologous antigen inhibited the binding to either P peptide or DNA coated onto the solid phase. These results show that sera in which the two specificities coexist contain antibodies endowed with a double binding ability for DNA and the P peptide.
Atorvastatin suppresses interferon-γ-induced neopterin formation and tryptophan degradation in human peripheral blood mononuclear cells and in monocytic cell lines
Authors: NEURAUTER G , WIRLEITNER B , LAICH A , SCHENNACH H , WEISS G , FUCHS D
Published: March 2003

Inhibitors of 3-hydroxy-3methylglutaryl-co-enzyme A (HMG-CoA) reductase, so-called statins, are used in medical practice because of their lipid-lowering effect and to reduce the risk of coronary heart disease. Recent findings indicate that statins also have anti-inflammatory properties and can modulate the immune response. In vitro, we investigated the effect of atorvastatin on the T cell/macrophage system in peripheral blood mononuclear cells (PBMC) and in the human monocytic cell lines THP-1 and MonoMac6. We monitored neopterin production and tryptophan degradation in PBMC after treatment with 10 µm and 100 µm atorvastatin in the presence or absence of 100 U/ml IFN-γ, 10 µg/ml phytohaemagglutinin (PHA) or 10 µg/ml concanavalin A (ConA) and in monocytic cell lines THP-1 and MonoMac6 with or without stimulation with 100 U/ml IFN-γ or 10 ng/ml to 1 µg/ml lipopolysaccharide (LPS). In stimulated PBMC 100 µm atorvastatin inhibited neopterin formation and tryptophan degradation completely, whereas 10 µm atorvastatin was only partially effective. Also in monocytic cell lines THP-1 and MonoMac6, atorvastatin was able to suppress IFN-γ- and LPS-induced formation of neopterin and degradation of tryptophan. Our data from PBMC agree well with previous investigations that statins inhibit T cell activation within the cellular immune response. In addition we demonstrate that atorvastatin directly inhibits IFN-γ-mediated pathways in monocytic cells, suggesting that both immunoreactivity of T cells and of monocyte-derived macrophages are down-regulated by this statin.
Antigen inhibition of collagen-induced arthritis is associated with up-regulation of IL-4 mRNA and induction of Ox40 on T cells in draining lymph nodes
Authors: MATTSSON L , LUNDBERG K , MÜSSENER E , JANSSON A , ERLANDSSON HARRIS H , LARSSON P
Published: March 2003

The addition of a foreign antigen to an inoculum completely inhibits the development of collagen-induced arthritis (CIA). However, the mechanism of this phenomenon, antigen -inhibition, is incompletely understood. Previous studies have demonstrated that the inhibition of arthritis is not mediated through suppression of the antibody response to cartilage antigens. In this paper we investigated cytokine mRNA levels in lymph nodes cells recovered 3, 7 or 16 days from animals immunized with either collagen II in IFA or OVA + collagen II in IFA. At day 7, but not at other time-points, IL-4 mRNA was up-regulated in the lymph nodes of OVA-inhibited non-arthritic animals compared to control animals which all developed arthritis. No significant differences between the two groups could be detected when expression of IFN-γ, IL-2, TNF-α, IL-1β or IL-10 mRNA was analysed. Flow cytometry analysis of draining lymph node cells demonstrated that the T cell marker Ox40 was up-regulated in the OVA-inhibited group. Our results indicate that the complete inhibition of CIA caused by addition of OVA to the collagen II inoculum is due to the presence of a TH2 environment resulting from an increased production of IL-4 mRNA and a parallel increase in Ox40+ T cells.
Role of interleukin-12 in the regulation of CD4+ T cell apoptosis in a mouse model of asthma
Authors: KODAMA T , KURIBAYASHI K , NAKAMURA H , FUJITA M , FUJITA T , TAKEDA K , DAKHAMA A , GELFAND E W , MATSUYAMA T , KITADA O
Published: March 2003

Allergic asthma, a chronic inflammatory disease of the airways, is characterized by the presence of T helper 2 cells and eosinophils in sputum, bronchoalveolar lavage, and mucosal biopsy specimens. Although the T helper 1-promoting cytokine, interleukin-12, is capable of inhibiting the T helper 2-driven asthma symptoms and bronchial responsiveness, the specific mechanisms underlying these interleukin-12 actions are unclear. The anti-allergic response to interleukin-12 is only partially dependent on interferon-γ, which induces apoptosis by enhancing expression of Fas antigen. We therefore investigated in vivo whether the anti-allergic action of interleukin-12 is mediated through induction of apoptosis. C57BL/6 mice immunized to ovalbumin by intraperitoneal injection were challenged three times with an ovalbumin aerosol every second day for 7 days. Recombinant interleukin-12 was administered intravenously after the final challenge. After the last ovalbumin challenge, mice were examined for effects of interleukin-12 on inflammatory cell infiltration and apoptosis in the lung as detected by terminal deoxynucleotidyl transferase-mediated deoxyribonucleoside triphosphate nick end-labelling.

Administration of interleukin-12 reduced ovalbumin-induced pulmonary eosinophilia (P < 0·01) and CD4+ T cell infiltration (P < 0·01). Moreover, treatment with interleukin-12 shortly after ovalbumin inhalation resulted in both increased interferon-γ production (P < 0·01) and enhanced apoptosis of CD4+ T cells in allergic airway infiltrates (P < 0·05). These results suggest that the beneficial effects of interleukin-12 in asthma may include enhancement of apoptosis of CD4+ T cells in airways.
Dendritic cells infected with adenovirus expressing the thyrotrophin receptor induce Graves’ hyperthyroidism in BALB/c mice
Authors: KITA-FURUYAMA M , NAGAYAMA Y , PICHURIN P , MCLACHLAN S M , RAPOPORT B , EGUCHI K
Published: March 2003

Dendritic cells (DCs) are the most potent antigen-presenting cells and a prerequisite for the initiation of primary immune response. This study was performed to investigate the contribution of DCs to the initiation of Graves’ hyperthyroidism, an organ-specific autoimmune disease in which the thyrotrophin receptor (TSHR) is the major autoantigen. DCs were prepared from bone marrow precursor cells of BALB/c mice by culturing with granulocyte macrophage-colony stimulating factor and interleukin−4. Subcutaneous injections of DCs infected with recombinant adenovirus expressing the TSHR (but not β-galactosidase) in syngeneic female mice induced Graves’-like hyperthyroidism (8 and 35% of mice after two and three injections, respectively) characterized by stimulating TSHR antibodies, elevated serum thyroxine levels and diffuse hyperplasitc goiter. TSHR antibodies determined by ELISA were of both IgG1 (Th2-type) and IgG2a (Th1-type) subclasses, and splenocytes from immunized mice secreted interferon-γ (a Th1 cytokine), not interleukin-4 (a Th2 cytokine), in response to TSHR antigen. Surprisingly, IFN-γ secretion, and induction of antibodies and disease were almost completely suppressed by co-administration of alum/pertussis toxin, a Th2-dominant adjuvant, whereas polyriboinosinic polyribocytidylic acid, a Th1-inducer, enhanced splenocyte secretion of IFN-γ without changing disease incidence. These observations demonstrate that DCs efficiently present the TSHR to naive T cells to induce TSHR antibodies and Graves’-like hyperthyroidism in mice. In addition, our results challenge the previous concept of Th2 dominance in Graves’ hyperthyroidism and provide support for the role of Th1 immune response in disease pathogenesis.
Circulating endothelial cells in Kawasaki disease
Authors: NAKATANI K , TAKESHITA S , TSUJIMOTO H , KAWAMURA Y , TOKUTOMI T , SEKINE I
Published: March 2003

summary

Recent reports have demonstrated that circulating endothelial cells (CECs) are observed in several diseases with vascular injury. Because Kawasaki disease (KD) is one type of systemic vasculitis, we hypothesized that an increased number of CECs may be associated with the appearance of complicated coronary artery lesions (CAL). In the present study we investigated the enumeration and origin of CECs in 20 patients with KD, using an immunohistochemical method with monoclonal antibodies: clone P1H12 against ECs and clone AC133 against endothelial progenitor cells (EPCs), which were derived from the bone marrow. The mean number of CECs increased significantly (P < 0·05) from the acute through the subacute phases of KD compared with both the convalescent phase of KD and healthy children. The mean number of CECs was significantly (P < 0·05) higher in six KD patients with CAL than in 14 KD patients without CAL. The population of EPCs in the total CECs in KD was 4·4 ± 1·2% (range 0–18%). The number of EPCs during the subacute phase was also significantly higher (P < 0·05) in KD patients with CAL than in those without CAL. Our findings indicate that the number of CECs increase in KD vasculitis and suggest that the increased numbers of CECs and EPCs may reflect the EC damage of this disease.
Oxygen free radicals and systemic autoimmunity
Authors: AHSAN H , ALI A , ALI R
Published: March 2003

Reactive oxygen species generated during various metabolic and biochemical reactions have multifarious effects that include oxidative damage to DNA leading to various human degenerative and autoimmune diseases. The highly reactive hydroxy radical (·OH) can interact with chromatin and result in a wide range of sugar and base-derived products, DNA–protein cross-links and strand breaks. Studies from our laboratory have demonstrated that after modification the DNA becomes highly immunogenic and the induced antibodies exhibit variable antigen-binding characteristics. Systemic lupus erythematosus, a prototype autoimmune disease, is characterized by the presence of autoantibodies to multiple nuclear antigens. The detection of 8-hydroxyguanosine in the immune complex derived DNA of systemic lupus erythematosus patients reinforces the evidence that reactive oxygen species may be involved in its pathogenesis. Increased apoptosis and decreased clearance of apoptotic cells as observed in systemic lupus erythematosus (SLE) might well be a contributory factor in systemic autoimmunity. Clinically, titres of autoantibodies are closely related to the degree of renal inflammation. Anti-DNA antibodies may combine with circulating antigen and contribute to the deposition of immune complexes in renal glomeruli.
Combination therapy with GH and cyproterone acetate does not improve final height in boys with non‐GH‐deficient short stature
Authors: Kawai Masahiko , Momoi Toru , Yorifuji Tohru , Muroi Junko , Yamanaka Chutaro , Sasaki Hiroshi , Furusho Kenshi
Published: March 2003

BACKGROUND AND OBJECTIVE

Recently, we reported that GH therapy without gonadal suppression (GS) decreased the final height of boys with non‐GH‐ deficient short stature by decreasing the height standard deviation score (SDS) for bone age (BA) during puberty. Combination therapy with GH and GS has been reported to suppress bone maturation and improve final height in some cases. We evaluated the effects of combination therapy with GH and GS using cyproterone acetate on the final height of boys with non‐GH‐deficient short stature.

PATIENTS

Fifty nine boys with non‐GH deficient short stature were observed retrospectively until they reached their final height. The boys were divided into 3 groups: Group A consisted of 26 boys who were not treated with GH, group B consisted of 13 boys who were treated with GH alone, and group C consisted of 20 boys who were treated with combination therapy with GH and GS using cyproterone acetate. At the start of observation, the height SDS for BA and projected height were matched among these three groups.

RESULTS

The mean ± SDS of the final height for groups A, B, and C were 162.7 ± 5.3 cm, 155.4 ± 4.9 cm, and 161.9 ± 3.2 cm, respectively. GH therapy did not affect the height SDS for BA during the prepubertal period. GH therapy without GS decreased the height SDS for BA during puberty in group B. Combination therapy with GH and cyproterone acetate increased the height SDS for BA between 12 and 14 years BA in group C. However, after GS therapy was discontinued at 14 years BA, the height SDS for BA gradually decreased and eventually reached the same value as that in group A.

CONCLUSIONS

GH therapy during the prepubertal period did not improve the final height of boys with non‐GH‐deficient short stature. GH therapy without GS decreased pubertal height gain, resulting in reduced final height. Combination therapy with GH and GS using cyproterone acetate decelerated the bone maturation during puberty which might be accelerated by GH therapy, but did not improve the final height which might have been attained without treatment.
Aromatase in the human central nervous system
Authors: Sasano Hironobu , Takahashi Kazuhiro , Satoh Fumitoshi , Nagura Hiroshi , Harada Nubuhiro
Published: March 2003

OBJECTIVE

Oestrogen produced locally by aromatase is thought to participate in numerous biological functions in the adult central nervous system (CNS). However, little is known about aromatase expression in the human CNS.

DESIGN

We examined aromatase expression in human brain regions, (4 men, 2 women) obtained from autopsy, by reverse transcriptase (RT)‐polymerase chain reaction (PCR) and also studied alternative use of multiple exons 1 of its gene, which is involved in tissue specific expression of aromatase in human.

RESULTS

The amount of aromatase mRNA determine by RT‐PCR assay in 6 cases tended to be highest in pons, thalamus, hypothalamus and hippocampus. Analysis of multiple exons 1 revealed that 1f, considered specific for brain, as well as 1b (fibrolast type) and 1d (gonadal type), were expressed. 1d and 1f tended to be utilized in hypothalamus, thalamus and amygdala. The amount of overall mRNA expression was also higher in hypothalamus, thalamus and amygdala than in other regions of the brain. There were no differences of utilization of exons 1 and mRNA expression of aromatase between female and male brain.

CONCLUSIONS

These results demonstrate that aromatase is expressed widely in various regions of human brain tissues in both men and women.
A randomized trial of short‐term treatment of Graves' disease with high‐dose carbimazole plus thyroxine versus low‐dose carbimazole
Authors: Grebe Stefan K. G. , Feek Colin M. , Ford Henry C. , Fagerström Jocelyn N. , Cordwell Diane P. , Delahunt John W. , Toomath Robyn J.
Published: March 2003

OBJECTIVE

The optimal treatment regimen with thionamide drugs remains a matter for debate. We have investigated whether high doses of carbimazole, when compared with low doses, reduce relapse rates of Graves' disease.

DESIGN

In an open label, randomized, prospective trial of treatment of Graves' disease we compared high doses of carbimazole (6 months of 100 mg carbimazole per day plus thyroxine) to low‐dose carbimazole treatment (starting at 25 mg and titrating the carbimazole dose with the aim to maintain serum thyroid function test results within the normal reference range).

PATIENTS

Thirty‐seven patients with a first episode of Graves' disease were enrolled.

MEASUREMENTS

During the 6 months of treatment we evaluated the rate of normalization of serum thyroid function tests, changes in serum thyroid auto‐antibody levels and the rate of side‐effects during treatment. After completion of the 6‐month treatment course patients were observed for 2 years for evidence of relapse of Graves' disease.

RESULTS

There were no differences between the two groups either in the rate of normalization of serum thyroid function tests or in serum thyroid auto‐antibody levels during treatment. Of the 17 patients randomized to high‐dose treatment seven suffered treatment side‐effects, compared to only one of the 20 patients receiving low‐dose treatment (P < 0.006). There was no significant difference in 2‐year post‐treatment remission rates on an intention‐to‐treat basis between the two treatment groups (18.7% vs. 5.9%, P = NS). However, for those patients who completed 6 months of treatment (high‐dose group = 9, low‐dose group = 16), multivariate survival analysis demonstrated a significantly longer median relapse‐free interval (P < 0.04) in the high‐dose group (27 weeks; 25th percentile: 9.6 weeks, 75th percentile: 75 weeks) versus the low‐dose group (6 weeks; 25th percentile: 4.8 weeks, 75th percentile: 13.1 weeks).

CONCLUSIONS

High‐dose carbimazole treatment delays, but does not prevent, relapse from Graves' disease in those patients able to tolerate the treatment. However, it leads to more frequent side‐effects than conventional dose treatment.
Lipoprotein lipase mass and activity in post‐heparin plasma from subjects with intra‐abdominal visceral fat accumulation
Authors: Kobayashi Junji , Tashiro Jun , Murano Shunichi , Morisaki Nobuhiro , Saito Yasushi
Published: March 2003

OBJECTIVES

The purpose of this study was to investigate the possibility of impaired lipolysis of triglyceride‐rich lipoproteins in patients with abdominal visceral fat accumulation by assessing two major lipolytic enzymes in the plasma, lipoprotein lipase (LPL) and hepatic lipase (HL).

DESIGN AND PATIENTS

A total of 31 patients [20 men, 11 women, age 50 ± 7 years old, body mass index (BMI) 26 ± 2 kg/m2 (mean ± sd)] were analyzed. Visceral fat and subcutaneous fat areas were evaluated using a computerized tomographic (CT) method at the level of the umbilicus. Total lipolytic activity in the postheparin plasma (PHP) was measured using Triton X‐100‐emulsified triolein and LPL activity was calculated as the activity in whole plasma inhibited by the 5D2 monoclonal antibody for LPL. LPL enzyme mass was determined by a sandwich enzyme immunoassay.

RESULTS

The visceral fat area was found to be negatively correlated with LPL mass (V vs LPL mass, r = − 0.37, P = 0.04) in PHP and had a tendency toward negative correlation with the LPL activity in the PHP (V vs LPL activity, r = − 0.29, P = 0.12). Subcutaneous fat area, on the other hand, did not show any correlation with LPL activity (r = 0.13, P = 0.49) or mass (r = 0.22, P = 0.25) in the PHP. The visceral fat area was found to be positively correlated with fasting serum insulin levels (r = 0.67, P < 0.01). Body mass index (BMI) was not correlated with LPL mass or activity in the PHP. Multi‐regressional analysis showed that abdominal visceral fat could be correlated with LPL mass in the PHP, independently of fasting serum insulin. The HL activity from PHP of the patients did not show significant correlation with visceral fat area, subcutaneous fat area or body mass index.

CONCLUSIONS

Fat distribution affects LPL mass and activity, either directly or via another metabolic abnormality such as insulin resistance, leading to impaired hydrolysis of triglycerides in chylomicrons and very low density lipoproteins (VLDL) in these subjects.
Fibromyalgia — a syndrome associated with decreased nocturnal melatonin secretion
Authors: Wikner Johan , Hirsch Ulf , Wetterberg Lennart , Röjdmark Sven
Published: March 2003

OBJECTIVE

Most patients with fibromyalgic syndrome (FMS) complain of sleep disturbances, fatigue, and pain. These symptoms might be a consequence of changed melatonin (MT) secretion, since MT is known to have sleep promoting properties. Moreover, serum concentrations of two MT precursors (tryptophan and serotonin) — affecting both sleep and pain perception — appear to be low in patients with FMS. Therefore, the objective of this investigation was to study whether serum MT (s‐MT) level is also low in these patients.

DESIGN

Eight patients with FMS and 8 healthy sex‐, BMI‐, and age‐matched controls were included in the study. s‐MT concentrations were determined every second hour between 1800 and 0800 h. Urine was collected between 2200 and 0700 h for determination of urinary MT excretion. To evaluate total MT secretion between 1800 and 0800 h and MT secretion during the hours of darkness (between 23 and 07 h) individual MT areas under the curve (AUC) were calculated and expressed as group means.

RESULTS

The FMS patients had a 31% lower MT secretion than healthy subjects during the hours of darkness (MT AUC 2300–0700 h (mean ± SEM): 1.70 ± 0.17 vs 2.48 ± 0.38 nmol/l; P < 0.05). Also the s‐MT peak value was significantly lower in the patient group: 0.28 ± 0.03 vs 0.44 ± 0.06 nmol/l; P < 0.05).

CONCLUSION

Patients with fibromyalgic syndrome have a lower melatonin secretion during the hours of darkness than healthy subjects. This may contribute to impaired sleep at night, fatigue during the day, and changed pain perception.
Reproducibility of the low dose dexamethasone suppression test: comparison between direct plasma and salivary cortisol assays
Authors: Reynolds  , Bendall  , Whorwood  , Wood  , Walker  , Phillips
Published: March 2003

BACKGROUND

The low dose dexamethasone suppression test (DST) has been used to detect subtle variations in the feedback suppression of the hypothalamic–pituitary–adrenal axis, which may contribute to the pathogenesis of several diseases including depression, the metabolic syndrome and coronary artery disease. Little is known about the reproducibility of this test, or whether the test can be combined with analysis of salivary cortisol which would offer a significant advantage over plasma in population studies.

SUBJECTS AND DESIGN

A low dose DST was carried out in 29 healthy subjects (14 men, 15 women), aged 24–54 (mean 35.1) years, on two separate occasions 1–10 weeks apart. Following the administration of 0.25 mg dexamethasone (DXM) at 2200 h, plasma and saliva were sampled at 0830 h the next day. Cortisol was measured by radioimmunoassay in plasma and time‐resolved immunofluorescent assay (‘DELFIA’) in saliva. Bland–Altman plots were produced for post‐DXM plasma and salivary cortisol measures and used to derive a coefficient of repeatability for each measure, which describes the range of cortisol measurements within which 95% of repeated measurements will fall.

RESULTS

The baseline, pre‐DXM cortisol concentrations were far more variable for saliva (mean 16.5, range 4.4–34 nmol/l) than for plasma (mean 407.5; range 232–958 nmol/l). Following DXM both measurements showed an approximately 30% suppression from baseline but the variability of salivary cortisol was much greater. From the Bland–Altman plots the 95% range for the differences about their mean was calculated and used as an indication of repeatability. For plasma 95% of differences were within 0.78 log units, indicating that a repeated measurement was approximately half as small or twice as large as the first. For saliva 95% of differences were within 1.64, indicating that a repeated measurement was approximately five times as small or five times as large as the first.

CONCLUSIONS

Assessment of dexamethasone suppression by salivary cortisol measurement is far less repeatable than the use of plasma cortisol. In the context of field studies of dexamethasone suppression, salivary cortisol measurements may only be appropriate for large numbers of subjects.
Is opioidergic activity responsible for the circadian variation observed in the gonadotrophin responsiveness of early follicular phase women?
Authors: Rossmanith Winfried G. , Grasshof Claudia
Published: March 2003

OBJECTIVES

In women, the gonadotrophin response to gonadotrophin‐releasing hormone (GnRH) displays a circadian rhythm during the early follicular phase (EFP), with GnRH‐stimulated luteinizing hormone (LH) and follicle‐stimulating hormone (FSH) release found to be markedly decreased at night. Since the opioidergic inhibition of gonadotrophin secretion is selectively enhanced at night, we reasoned that the circadian changes in the gonadotrophin responsiveness to GnRH might be related to a nocturnal increase of opioidergic activity.

STUDY DESIGN

Eleven women with normal menstrual cycles were studied in the EFP on four different occasions in random order. Studies were conducted either during the day (0900–1300 h) or at night (2100–0100 h). During these times, GnRH (25 μg i.v.) was administered in conjunction with either saline (as control) or naloxone (4 mg i.v.).

MEASUREMENTS

Frequent blood samples were obtained before and after GnRH stimulation for determination of basal sex steroid and gonadotrophin concentrations by immunoradiometric assays.

RESULTS

While oestradiol levels were comparable (P  > 0.3) at all times, progesterone concentrations were significantly (P < 0.01) higher during day than during night hours, with no difference between control and naloxone conditions. Gonadotropin responses to GnRH stimulation were not significantly different between day and night times, nor did they vary between control and naloxone conditions.

CONCLUSIONS

Opioidergic blockade imposed by naloxone did not noticeably change GnRH‐stimulated gonadotrophin release at any of the study times. We therefore infer that mechanisms other than a nocturnal increase of opioidergic inhibition may account for eventual circadian changes in the gonadotrophin responsiveness of early follicular phase women.
The use of β‐subunits of gonadotrophin hormones in the follow‐up of clinically non‐functioning pituitary tumours
Authors: Greenman Yona , Tordjman Karen , Sömjen Dalia , Reider‐Groswasser Irith , Kohen Fortune , Ouaknine George , Stern Naftali
Published: March 2003

OBJECTIVE

Clinically nonfunctioning pituitary adenomas (NFA) are mostly of gonadotroph origin. However, increased levels of circulating hormones or subunits in patients with NFA usually do not cause clinical symptoms, nor are they used as biological tumour markers. In this study we assessed the value of measuring β subunits of gonadotrophin hormones in the post‐operative follow‐up of patients bearing these tumours.

DESIGN

Patients harbouring NFA were studied before and three months after transphenoidal pituitary surgery. β‐LH and β‐FSH levels were measured before and following TRH administration on the two occasions. Hormone levels were analyzed in relation to imaging studies performed before and after surgery.

PATIENTS

Twenty four patients operated at the Tel Aviv‐Sourasky Medical Centre for NFA.

RESULTS

Pathological β‐FSH and β‐LH levels were detected in 79% and 60% of patients respectively. β‐LH levels decreased after surgery but there were no significant changes in β‐FSH levels. There was a tendency for tumours with high basal β‐LH levels to be larger and to have a poor surgical outcome. Normalization of β‐LH levels post‐operatively was usually associated with a decrease in tumour mass or complete removal of the tumour. Persistent pathological responses of β‐LH to TRH after surgery were common in patients with residual tumours on imaging. Nevertheless there were exceptions to this pattern, rendering post‐operative β‐LH levels insufficiently reliable as a marker for the presence of residual tumour.

CONCLUSION

Although there appears to be a relationship between β‐LH levels, tumour size and surgical outcome, this association is presently insufficient to allow the routine use of either basal or TRH induced β‐LH responses in the post‐surgical follow‐up of clinically nonfunctioning pituitary adenomas.
Leptin and the pituitary–thyroid axis: a comparative study in lean, obese, hypothyroid and hyperthyroid subjects
Authors: Pinkney Jonathan H. , Goodrick Steven J. , Katz Jonathan , Johnson Andrew B. , Lightman Stafford L. , Coppack Simon W. , Mohamed‐Ali Vidya
Published: March 2003

OBJECTIVE

To study interactions between leptin and the pituitary–thyroid axis, both in euthyroid and dysthyroid states.

SUBJECTS AND MEASUREMENTS

We investigated the relationships of plasma leptin to levels of free thyroid hormones and TSH in 18 patients with newly diagnosed hyperthyroidism, 22 with newly diagnosed primary hypothyroidism, and 32 lean (body mass index [BMI] < 30) and 37 obese (BMI > 30 kg/m2) euthyroid subjects. Hypothyroid patients were restudied during thyroxine replacement treatment.

RESULTS

Median [interquartile range] plasma leptin concentrations were highest in obese euthyroid subjects (31.5 [19.0–48.0] and in untreated hypothyroid patients (19.2 [11.5–31.5]), and lowest levels in untreated hyperthyroid patients (8.9 [5.5–11.1]) and lean euthyroid control subjects (6.6 [3.9–14.4] μg/l (Kruskall–Wallis one‐way analysis of variance; P < 0.0001). In euthyroid subjects, plasma leptin levels were higher in obese than in lean subjects (P < 0.00001). In obese subjects plasma levels of TSH correlated with percentage body fat (r = 0.67; P < 0.001) and plasma leptin (r = 0.61; P < 0.001). In untreated hyperthyroid subjects plasma leptin was unrelated to free T3, and in untreated hypothyroidism plasma leptin was unrelated to either free T3 or TSH concentrations (all P = NS). In untreated hyperthyroid, but not hypothyroid, patients plasma leptin concentrations correlated with BMI (r = 0.57; P = 0.02). Treatment of hypothyroidism with thyroxine resulted in a significant reduction in plasma leptin concentrations from 20.8 (11.8 to 31.6) to 12.9 (4.6–21.2) μg/l (P = 0.005), but BMI did not change significantly in the hypothyroid subjects being studied prospectively.

CONCLUSIONS

(i) In euthyroid subjects, plasma leptin and TSH levels correlate, and both are positively correlated with adiposity. (ii) Plasma leptin was significantly elevated in hypothyroid subjects, to levels similar to those seen in obese euthyroid subjects. (iii) Treatment of hypothyroidism resulted in a reduction in the raised plasma leptin levels. The data are consistent with the hypothesis that leptin and the pituitary–thyroid axis interact in the euthyroid state, and that hypothyroidism reversibly increases leptin concentrations.
β3‐adrenoreceptor gene polymorphism and leptin. Lack of relationship in type 2 diabetic patients
Authors: Vendrell  , Gutierrez  , Broch  , Fernandez‐Real  , Aguilar  , Richart
Published: March 2003

INTRODUCTION

The β3‐Adrenergic receptor (β3AR) and leptin are molecules involved in the regulation of energy balance. Recently, a mutation in the β3AR gene (Trp64Arg) has been reported to be associated with features of insulin resistance, weight gain and early onset of Type 2 Diabetes Mellitus. The aim of this study was to determine the frequency and clinical characteristics of the Trp64Arg mutation in the β3AR gene in Type 2 diabetic patients, its relationship with leptin levels, and its role in microangiopathic complications.

SUBJECTS AND METHODS

187 Type 2 diabetic patients and 100 unrelated non‐diabetic subjects were studied. There was no difference between the diabetic and nondiabetic subjects in the frequency of the Trp64 and Arg64 alleles (92.5% vs. 92.3% and 7.5% vs. 7.7%, respectively). Type 2 diabetic patients were divided into two groups according to the presence (n = 27) or absence of the mutation (n = 160).

RESULTS

Mutation of the β3AR gene was not associated with any differences either in the clinical and metabolic parameters or microangiopathic complications. Type 2 diabetic patients carrying the Arg64 allele tended to have a lower diabetes duration, but this was not statistically significant. Plasma leptin levels were not different according to the β3AR genotype.

CONCLUSIONS

The Trp64Arg mutation is not a major determinant of Type 2 diabetes and its microangiopathic complications. Moreover, this mutation was not clinically relevant in leptin regulation.
Secondary adrenal insufficiency associated with autoimmune disorders: a report of twenty‐five cases
Authors: Kasperlik‐Załuska A. A. , Czarnocka B. , Czech W. , Walecki J. , Makowska A. M. , Brzeziński J. , Aniszewski J.
Published: March 2003

OBJECTIVE

Addison's disease is frequently a component of autoimmune polyendocrinopathies while secondary adrenal insufficiency associated with autoimmune disorders is believed to be a rare event. We present a series of patients with secondary adrenal insufficiency coexisting with autoimmune diseases and/or antithyroid autoantibodies.

DESIGN AND PATIENTS

Among a group of 102 patients with secondary adrenal failure of unknown origin diagnosed at the Department of Endocrinology of the Centre for Postgraduate Medical Education (Warsaw, Poland) we have identified a group with associated autoimmune disorders. Thyroid abnormalities occurred most frequently. Other diseases included insulin‐dependent diabetes mellitus, pernicious anaemia, vitiligo, premature ovarian failure and autoimmune thrombocytopaenia. There were 23 women and one man aged 17–72 years at the time of investigation. Additionally, we included a woman with Addison's disease in whom the ACTH deficiency appeared 18 years after the onset of primary adrenal hypofunction.

MEASUREMENTS

Pituitary–adrenal function tests comprised urinary excretion of 17‐hydroxycorticosteroids in basal conditions and during a 2‐day tetracosactrin test, plasma concentrations of ACTH and cortisol, and a 2‐day metyrapone test (in eight cases). Thyroid function and immunity tests were: TSH, thyroxine, the antithyroglobulin, antimicrosomal and anti‐peroxidase autoantibodies. Other endocrine studies included: serum LH, FSH and PRL.

RESULTS

The 17‐hydroxycorticosteroid values, both basally and during stimulation tests were consistent with a diagnosis of secondary adrenal insufficiency. Serum cortisol and plasma ACTH concentrations were low. In 14 patients primary hypothyroidism was confirmed by low T4 levels. In three patients subclinical primary hypothyroidism was revealed (elevated TSH levels). Three patients who had a past history of Graves' disease were euthyroid at the time of investigation. Twenty‐three patients had antibodies against peroxidase. Most patients had gonadotrophins and PRL values within normal limits.

CONCLUSIONS

The co‐existence of autoimmune disorders with secondary adrenal insufficiency suggests an autoimmune aetiology for the ACTH deficiency.
Different immunohistochemical patterns of TGF‐β1 expression in benign and malignant adrenocortical tumours
Authors: Boccuzzi Adriana , Terzolo Massimo , Cappia Susanna , De Giuli Paolo , De Risi Carolina , Leonardo Eugenio , Bovio Silvia , Borriero Mirella , Paccotti Piero , Angeli Alberto
Published: March 2003

OBJECTIVE

Transforming growth‐factor β1 (TGF‐β1) influences a number of specific functions of adrenocortical cells in several animal species. The aim of our study was to evaluate by immunohistochemical analysis the presence and distribution of TGF‐β1 in normal adrenal tissue and in different adrenal tumours.

PATIENTS

We analysed 8 functioning (5 adenomas and 3 carcinomas) and 15 non functioning (6 adenomas and 9 carcinomas) adrenal tumours and 6 normal adrenal glands.

RESULTS

In normal adrenal glands, the glomerulosa and the reticularis zones displayed diffuse cytoplasmic staining, while the fasciculata zone was almost completely negative. Functioning adenomas displayed cytoplasmic staining restricted to compact cells while in nonfunctioning adenomas, prevalently composed by clear cells, no staining was observed. Overall, adrenal carcinomas were characterized by the lack of cytoplasmic positivity and by sporadic positive cells around vessels both in functioning and in nonfunctioning tumours.

CONCLUSIONS

TGF‐β1 expression is associated with active steroid secretion in normal adrenal tissue, as well as in benign cortical adenomas, while this relationship is lost in primary adrenal malignancies. These data provide indirect evidence for a regulatory role played by TGF‐β1 on steroid secretory pathways.
Kallmann's syndrome: is it always for life?
Authors: Quinton  , Cheow  , Tymms  , Bouloux  , Wu  , Jacobs
Published: March 2003

OBJECTIVE

Kallmann's syndrome (KS) is defined by the association of olfactory deficit with irreversible, congenital gonadotrophin deficiency (IHH). We present evidence for the existence of a variant form of KS, in which endogenous gonadotrophin secretion recovers spontaneously in later life.

DESIGN

Longitudinal clinical study.

PATIENTS

Five men with anosmia or severe hyposmia, who originally presented in their late teens or early twenties as a result of severe pubertal delay and were thus presumed to have KS.

RESULTS

Spontaneous onset of endogenous gonadotrophin secretion, evidenced by progressive normalization of testicular volume and of serum testosterone concentration, occurred in these men over a period of years following the initial diagnosis.

CONCLUSIONS

This variant form of Kallman's syndrome is not well recognized and may well be under‐diagnosed. Once full virilization has been induced, males with congenital gonadotrophin deficiency whose testes have significantly increased in size should be reassessed, off androgen replacement therapy, to identify those who no longer require treatment.
Correlation between urinary cortisol and 6‐sulphatoxymelatonin rhythms in field studies of blind subjects
Authors: Skene  , Lockley  , James  , Arendt
Published: March 2003

OBJECTIVE

Blind individuals have different types of circadian rhythm disorders. In order to study these in the field reliable markers of circadian phase are required. The aim of the present study was to determine the usefulness of urinary cortisol as a marker rhythm in field studies. This was assessed by investigating the relationship between the cortisol rhythms and the previously determined melatonin rhythms from a large sample of blind people with different types of circadian rhythm disorders.

DESIGN

Field study

SUBJECT

Registered blind subjects (n = 49) classified as having light perception or better (n = 19, 12 men, 7 women, aged 23–61 years) or having no conscious perception of light (n = 30, 24 men, 6 women, aged 19–72 years) were studied in their normal environment.

MEASUREMENT

Sequential 4‐hourly urine samples (plus an overnight sample) were collected for 48 h each week for 3–5 weeks. Urinary cortisol and 6‐sulphatoxymelatonin were measured by radioimmunoassay.

RESULT

Irrespective of the type of circadian rhythm (entrained or free‐running), there was a significant correlation between the characteristics of the 6‐sulphatoxymelatonin and cortisol rhythms in the blind subjects.

CONCLUSION

Urinary cortisol is recommended as a useful marker of circadian phase in field studies in addition to urinary 6‐sulphatoxymelatonin measurements.
An adult patient with severe hypercalcaemia and hypocalciuria due to a novel homozygous inactivating mutation of calcium‐sensing receptor
Authors: Chikatsu Noriko , Fukumoto Seiji , Suzawa Miyuki , Tanaka Yuji , Takeuchi Yasuhiro , Takeda Shu , Tamura Yasuhiro , Matsumoto Toshio , Fujita Toshiro
Published: March 2003

Inactivating mutations in the calcium‐sensing receptor (CaSR) cause familial hypocalciuric hypercalcaemia (FHH) and neonatal severe hyperparathyroidism (NSHPT). Earlier investigations showed patients with FHH are heterozygous, and NSHPT are homozygous for inactivating mutations. However, one adult patient with severe hypercalcaemia and hypocalciuria has been reported to have a homozygous inactivating mutation in CaSR (Pro39Ala). This suggested that mutant CaSR in this patient had some residual activity and hypercalcaemia was not so severe as to be fatal. However, the function of this mutant CaSR was not evaluated. In the present study, we describe a novel homozygous mutation in an adult patient with severe hypercalcaemia and hypocalciuria, and evaluate the function of the mutant CaSRs. The DNA sequence of CaSR gene was determined by direct sequencing of the polymerase chain reaction product. The function of mutant CaSR was analysed by creating mutant cDNAs by in vitro mutagenesis, transfection of mutant cDNAs into HEK293 cells and measuring intracellular ionized Ca in response to changes in extracellular Ca. A 26‐year‐old Japanese woman showed marked hypercalcaemia with an elevated parathyroid hormone (PTH) level. Her consanguineous parents had asymptomatic hypercalcaemia with relative hypocalciuria. The proband had a homozygous mutation at codon 27 of CaSR gene (CAA→CGA, Gln27Arg). Her parents were heterozygous for this mutation. EC50 for Ca of this mutant CaSR (Gln27Arg) was 4.9 m
m. EC50 of another mutant CaSR (Pro39Ala) whose homozygous mutation was discovered in an adult patient was 4.4 m
m. These EC50s were significantly higher than that of wild‐type CaSR (3.7 } 0.1 m
m), but were the lowest among the reported EC50s for inactivating mutations of CaSR. These results indicate that serum Ca and PTH levels are determined by residual function of mutant CaSR in patients with homozygous mutation in CaSR, and that patients having homozygous mutant CaSRs with mild dysfunction do not suffer from fatal hypercalcaemia in infancy and can survive into adulthood.
Adrenal and gonadal contributions to urinary excretion and plasma concentration of epitestosterone in men — effect of adrenal stimulation and implications for detection of testosterone abuse
Authors: Kicman Andrew T. , Coutts Sarah B. , Cowan David A. , Handelsman David J. , Howe Chris J. , Burring Surrinder , Wu Frederick C. W.
Published: March 2003

OBJECTIVE

The ratio of urinary testosterone (T) to epitestosterone (EpiT) is used to detect T abuse in sport. Also, plasma or urinary concentrations of EpiT have been measured to assess testicular steroidogenesis during hormonal male contraception. Further investigations are required to evaluate the relative contributions of the testis and adrenal to EpiT production. To this purpose, we have compared basal urinary EpiT glucuronide and plasma EpiT and the response to synthetic adrenocorticotrophic hormone (ACTH) stimulation between eugonadal and hypogonadal men.

DESIGN AND SUBJECTS

The basal urinary excretion rate of EpiT glucuronide was determined in 34 eugonadal men. Six men, clinically diagnosed as hypogonadal, and 6 out of the 34 eugonadal men previously described, received an intramuscular injection of synthetic ACTH depot (1 mg) at 0800 h on two consecutive days. Blood samples were collected prior to and then at 1.5, 8, 24, 25.5, 32 and 48 h with respect to the first administration (0 h). 24‐h urine specimens were collected from 0800 h on days 1 and 2 (baseline) and 3 and 4 (stimulation).

MEASUREMENTS

Plasma EpiT, T and cortisol were measured by RIA and urinary EpiT and T, following glucuronide hydrolysis, by gas chromatography‐mass spectrometry (extract combines aglycones with a minor amount of urinary free steroids).

RESULTS

Basal excretion rates of EpiT glucuronide in eugonadal men (range:62–751 nmol/24 h) were considerably greater than in hypogonadal men (range: 3–34 nmol/24 h). Mean basal plasma EpiT in eugonadal men (1.32 ± 0.08 nmol/l) were greater than in hypogonadal men (0.68 ± 0.04 nmol/l). In each group, synthetic ACTH stimulation increased plasma cortisol 4‐fold. In eugonadal men, plasma and urinary EpiT were unchanged whereas plasma and urinary T glucuronide decreased in response to ACTH. In hypogonadal patients, ACTH increased plasma and urinary EpiT while plasma T remained unchanged.

CONCLUSION

The testes are the major source of epitestosterone, the adrenal contribution being relatively modest. Following adrenal stimulation, urinary epitestosterone glucuronide increases considerably in hypogonadal men but this increase is masked in eugonadal men because testicular production is probably suppressed by the ACTH‐induced rise in cortisol. Activation of the adrenal cortex results in no change or only a small decrease in the urinary T/EpiT ratio in eugonadal men.
A novel mutation in the TPO gene in goitrous hypothyroid patients with iodide organification defect
Authors: Santos Cecilia L. S. , Bikker Hennie , Rego Katia G. M. , Nascimento Antonio C. , Tambascia Marcos , De Vijlder Jan J. M. , Medeiros‐Neto Geraldo
Published: March 2003

OBJECTIVE

To screen and subsequently sequence the TPO gene for mutations in patients with congenital goitre, hypothyroidism and evidence for an organification defect (positive perchlorate discharge test).

PATIENTS

We have studied seven hypothyroid and congenitally goitrous patients from three unrelated families.

DESIGN AND MEASUREMENTS

We have measured serum thyroid hormone levels, 131I uptake, serum TSH and serum Tg concentrations. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified genomic DNA was used to screen for mutations in the TPO gene.

RESULTS

DGGE identified the presence of two frameshift mutations: a GGCC duplication in exon 8 (homozygous in one family and heterozygous in the other family) and a heterozygous insertion of a single nucleotide (C) at position 2505‐2511 in exon 14. In addition, we have detected an alteration in exon 11, not yet described in the literature, derived from a single nucleotide substitution of a C to G at position 2008, altering the well‐conserved amino acid domain among the peroxidases superfamily. This mutation in exon 11 was present in two families that showed heterozygous mutation for exon 8 or for exon 14.

CONCLUSIONS

These results could support the hypothesis for a putative compound heterozygosity pattern in the affected patients. The altered phenotype (goitre and hypothyroidism since birth) seems justifiable in view of the possible inactivating character of this novel mutation in exon 11.