bluuebunny/crossref_metadata_2025 huggingface dataset:
Interferon-gamma (IFN-γ) regulates production of IL-10 and IL-12 in human herpesvirus-6 (HHV-6)-infected monocyte/macrophage lineage
Authors: LI C , GOODRICH J M , YANG X
Published: November 2003

To determine whether HHV-6 infection induces expression and production of IL-10 and IL-12 in monocytes/macrophages, and to explore the influence of IFN-γ on cytokine production in HHV-6-infected cells, expression and production of IL-10 and IL-12 were evaluated through reverse transcription-polymerase chain reaction (RT-PCR) and sandwich ELISA. HHV-6 infection induced the expression and the production of IL-10 and IL-12 in monocytes and THP-1 cells. Kinetic study showed that the expression of IL-12 mRNA decreased with accumulation of IL-10 mRNA. Expression and production of IL-12 were markedly increased when anti-human IL-10 MoAbs were added to the cultures, implying that endogenous IL-10 induced by HHV-6 inhibited IL-12 production. Addition of increasing concentrations of IFN-γ to the cultures of HHV-6-infected cells enhanced the expression of IL-12 gene, while the accumulation of IL-10 mRNA was down-regulated. Determination of protein levels of IL-10 and IL-12 by ELISA also showed that IFN-γ increased IL-12 and decreased IL-10 production. These results suggest that IFN-γ regulates the production of IL-10 and IL-12 at transcriptional level mainly through inhibiting endogenous IL-10 production in HHV-6-infected monocyte/macrophage lineage.
Expression of human–Torpedo hybrid acetylcholine receptor (AChR) for analysing the subunit specificity of antibodies in sera from patients with myasthenia gravis (MG)
Authors: LOUTRARI H , KOKLA A , TRAKAS N , TZARTOS S J
Published: November 2003

The nicotinic AChR, a pentamer composed of α2βγ(or ε)δ subunits, is the autoantigen in the human autoimmune disease MG. Anti-AChR antibodies in MG sera bind mainly to conformational epitopes, therefore determination of their specificities requires the use of intact AChR. Indirect antibody competition studies have suggested that most MG antibodies are inhibited from binding to AChR by MoAb to the main immunogenic region (MIR) on the α-subunits. More recently, based on the knowledge that MG antibodies show little detectable cross-reaction with Torpedo AChR, we have shown, using mouse–Torpedo hybrid AChR, that most MG antibodies that detectably cross-react with the mouse AChR bind to the α-subunit. To analyse the whole anti-AChR antibody repertoire in MG sera, we expressed on stably transfected fibroblasts a novel human α+Torpedoβγδ AChR and compared the antibody titres against human, Torpedo, and the hybrid AChR. Direct information was provided for the subunit specificity of several MoAbs and sera from 50 MG patients. On average, at least 48% of the anti-AChR antibodies in the sera were directed against the α-subunit. Interestingly, the anti-α-subunit antibodies predominated in low titre (0.6–7.4 nm) but not in high titre (10–386 nm) sera, where they comprised on average 68% versus 23% of the antibodies, respectively. Finally, the directly determined anti-α-subunit antibodies and the anti-MIR antibodies defined by antibody competition were significantly correlated, thus suggesting that at least a significant fraction of the anti-MIR antibodies in MG sera bind to the α-subunit.
Density of γ/δ+ T cells in the jejunal epithelium of patients with coeliac disease and dermatitis herpetiformis is increased with age
Authors: SAVILAHTI E , ÖRMÄLÄ T , ARATO A , HACSEK G , HOLM K , KLEMOLA T , NEMETH A , MÄKI M , REUNALA T
Published: November 2003

Increased density of γ/δ T cell receptor (TCR)+ intraepithelial lymphocytes is the only characteristic in the jejunum of patients with coeliac disease and dermatitis herpetiformis which is not normalized on a gluten-free diet. We explored the age-dependent changes in intraepithelial γ/δ and α/β TCR+ cells from 137 biopsies from patients with coeliac disease and dermatitis herpetiformis and from controls. Biopsy specimens from 100 patients with coeliac disease and dermatitis herpetiformis and from 37 controls were studied with an immunohistochemical method using MoAbs to T cell receptors and peroxidase staining. An increase in the density of intraepithelial γ/δ T cells above the mean +2 s.d. of the density in controls was present in 97 of 100 specimens from patients with coeliac disease and dermatitis herpetiformis. The density of γ/δ+ cells of patients with coeliac disease and dermatitis herpetiformis on a normal gluten-containing diet showed a positive correlation with age (r = 0.45, P < 0.0001). In controls, the density of γ/δ+ cells remained low throughout the age-range studies, from age 0.6–57 years. In controls, α/β+ cells increased with age (r = 0.57, P < 0.001). The increase in density of intraepithelial lymphocytes with age is in agreement with their thymus-independent character and local proliferation.
Chronic sinusitis refractory to standard management in patients with humoral immunodeficiencies
Authors: BUEHRING I , FRIEDRICH B , SCHAAF J , SCHMIDT H , AHRENS P , ZIELEN S
Published: November 2003

Chronic refractory sinusitis is a common feature in patients with primary immunodeficiencies. The efficacy of standard therapeutic strategies is questionable. In an open trial we evaluated the efficacy of azithromycin, N-acetylcysteine and topical intranasal beclomethasone (100 μg twice daily for 6 weeks) in 16 patients with primary immunodeficiencies (median age 13.5 years, range 5–32 years). All patients suffered from chronic sinusitis despite regular immunoglobulin replacement therapy every 3 weeks. Magnetic resonance imaging (MRI) scans were performed before and after 6 weeks of treatment to evaluate morphological changes in the paranasal sinuses. Nasal swabs and washings were taken for microbial analysis and measurement of inflammatory mediators (IL-8, tumour necrosis factor-alpha (TNF-α), eosinophilic cationic protein (ECP)) before and post therapy. Inflammatory mediators in nasal secretions were significantly elevated in patients: IL-8 median 2436 pg/ml (range 441–5435 pg/ml), TNF-α 37.3 pg/ml (3.75–524 pg/ml) and ECP 33 ng/ml (1.5–250 ng/ml) versus age-matched healthy controls: IL-8 median 212 pg/ml (99–825 pg/ml), TNF-α 3.77 pg/ml (2.8–10.2 pg/ml) and ECP 1.5 ng/ml (1.5–14.8 ng/ml) (P < 0.0001). Inflammation of the maxillary sinuses was confirmed by MRI scans in all patients, additionally infection of the ethmoidal and frontal sinuses was recorded in five patients. Bacterial growth appeared in 11 out of 16 cultures. In spite of therapy, no improvement in sinal inflammation visualized by MRI was achieved. Moreover, no significant decrease in pathogens and levels of inflammatory mediators could be detected (IL-8 1141 pg/ml, 426–4556 pg/ml; TNF-α 13.9 pg/ml, 4.1–291.6 pg/ml; ECP 32.3 ng/ml, 3.7–58.4 ng/ml). Our results demonstrate that conventional management of sinusitis is of little benefit in patients with chronic refractory sinusitis with an underlying immunodeficiency. More studies are needed to test antibiotic regimens, probably combined with surgical drainage and anti-inflammatory agents.
Antagonistic effects of IL-4 and interferon-gamma (IFN-γ) on inducible nitric oxide synthase expression in bovine macrophages exposed to Gram-positive bacteria
Authors: JUNGI T W , BRCIC M , SAGER H , DOBBELAERE D A E , FURGER A , RODITI I
Published: November 2003

Cytokine-mediated modulation of nitric oxide (NO) production by bacteria-stimulated bovine macrophages was studied. When Salmonella dublin, as a prototypic Gram-negative organism, was used, NO generation was barely enhanced by recombinant bovine and ovine IFN-γ, but was suppressed by IL-4. Salmonella dublin-induced NO generation was not influenced by a panel of nine other cytokines. The panel included IL-1, tumour necrosis factor (TNF) and IFN-α, which are active in a similar mouse macrophage model. The tested cytokines were either homologous or known to interact with bovine cytokine receptors. Recombinant bovine and ovine IFN-γ were the only cytokines which strongly enhanced NO synthesis by macrophages exposed to the Gram-positive organism, Listeria monocytogenes. Listeria-induced NO generation was strongly suppressed by recombinant human and bovine IL-4, but not by IL-10 and transforming-growth-factor-beta. Thus, two cytokines characterizing a Th1 and a Th2 response up- and down-regulate, respectively, bacteria-induced NO generation in bovine macrophages, whereas nine other cytokines had little activity in this regard. This modulation was reflected in changes in the steady state levels of mRNA coding for inducible nitric oxide synthase. Combinations of IFN-γ and IL-4 suggested that the relative proportion of these cytokines determined whether bacteria-induced NO generation was up- or down-regulated. At saturating IL-4 concentrations, stimulation of bacteria-induced NO generation in macrophages by T cell supernatants was solely dependent on IFN-γ. This was shown by antibody neutralization experiments and by a close correlation between the capacity of supernatants to stimulate NO generation and the IFN-γ content, as determined by immunoassay.
Cytoplasmic detection of a novel protein containing a nuclear localization sequence by human autoantibodies
Authors: GARCIA-LOZANO J R , GONZALEZ-ESCRIBANO M F , WICHMANN I , NUÑEZ-ROLDAN A
Published: November 2003

A great diversity of antibodies directed to cell proteins has been described in sera of patients with autoimmune diseases. Most of these sera recognize nuclear components, but some others are directed against cytoplasmic autoantigens. Some of the antibodies directed to cytoplasmic autoantigens are well characterized, such as anti-mitochrondial, anti-ribosomal, anti-microsomal and anti-Golgi complex autoantibodies, but the target of many others remains unknown. In the last 5 years we have selected 32 sera with a characteristic speckled cytoplasmic pattern in indirect immunofluorescence (IIF) assay among a total of more than 31 000 sera from patients with any kind of autoimmune manifestation who attend our Connective Tissue Disease Clinic. Using a human cDNA expression library, we have identified a new autoantibody specificity named RCD-8 in five of these sera, directed to one cytoplasmic autoantigen. Affinity-purified antibodies eluted from a positive clone reproduced the same IIF cytoplasmic staining pattern as native serum and reacted with one single band of 160 kD on an immunoblot of HeLa cell extract. The sequence was found homologous to an autoantigen recently reported named Ge-1, and contains a nuclear localization sequence (NLS), an active protein domain made by a contiguous stretch of amino acids which allows the selective entry of the protein into the nucleus. The five patients whose sera exhibited this new autoantibody specificity displayed different autoimmune pathological profiles.
Immune response to Plasmodium falciparum antigens in Cameroonian primigravidae: evolution after delivery and during second pregnancy
Authors: FIEVET N , COT M , RINGWALD P , BICKII J , DUBOIS B , LE HESRAN J Y , MIGOT F , DELORON P
Published: November 2003

Mechanisms responsible for the increase in malaria susceptibility during pregnancy, and in particular during the first pregnancy, have not been elucidated. T and B cell responses to leucoagglutinin, bacille Calmette–Guérin (BCG) and to six Plasmodium falciparum antigens were longitudinally investigated in 33 pregnant women during their first pregnancy, after delivery, and during second pregnancy. Parasitological data obtained from the same women during and after the first pregnancy demonstrated the higher risk of P. falciparum infection during this pregnancy. Plasma levels of antibodies to Pf155/RESA were lower during pregnancy than after delivery. Conversely, antibodies to P. falciparum asexual blood stages were higher during pregnancy than after delivery, suggesting that during pregnancy the regulation of antibody production may be variously impaired depending upon the antigens. The most striking finding of the present study is the impairment of the IL-2 cellular response during the first pregnancy. Conversely, proliferative responses, as well as IL-4 and interferon-gamma (IFN-γ) responses, were either unaffected or moderately enhanced. No difference in humoral and cellular responses was observed between first and second pregnancy. The impairment of the IL-2 responses involved the response to malaria peptides and proteins, as well as the response to non-malarial antigens and to the mitogen leucoagglutinin. Thus, the alteration of malaria immunity might rather fall into the general frame of the depression of cellular immunity during pregnancy than involve a specific malaria phenomenon.
Presence of glomerular basement membrane (GBM) antibodies in HIV− patients with Pneumocystis carinii pneumonia
Authors: CALDERON E J , WICHMANN I , VARELA J M , RESPALDIZA N , REGORDAN C , FERNANDEZ-ALONSO J , MEDRANO F J , CANO S , CUELLO J A , NUÑEZ-ROLDAN A
Published: November 2003

Following the unexpected finding of antibodies to GBM in a patient with Pneumocystis carinii pneumonia in the absence of kidney abnormalities, the presence of anti-GBM antibodies was analysed in 14 patients with pulmonary P. carinii infection who did not have clinical evidence of autoimmune glomerulonephritis. Patients were divided into three groups: HIV− with P. carinii pneumonia (n = 4), HIV+ with P. carinii pneumonia (n = 5) and HIV− carriers of P. carinii without pneumonia (n = 5). As control groups, HIV− patients with community-acquired non-P. carinii pneumonia (n = 6) and healthy individuals (n = 16) were included. Anti-GBM antibodies, studied with a quantitative enzyme immunoassay (EIA) for anti-α3 chain of collagen IV antibodies, were detected in three out of the four HIV− patients with P. carinii pneumonia, but not in any individuals of the other categories. These results suggest that P. carinii alveolar injury or the host response to the organism could affect the basal membrane Goodpasture antigen or a similar antigen, and induces anti-GBM antibody production in HIV− patients, and support the hypothesis that, at least in some cases, Goodpasture's syndrome could be triggered by an alveolar lesion induced by a P. carinii pneumonia.
Comparative study of the complement-activating and specific IgE-binding properties of ragweed pollen allergen
Authors: HIDVÉGI T , BERRENS L , VARGA L , MARAÑON F , SCHMIDT B , KIRSCHFINK M , FÜST G
Published: November 2003

Previous reports have defined the capacity of ragweed pollen extract (RWA) to activate human complement (C) in fluid phase through the classical pathway and have ascertained a strong correlation between the extent of complement activation and the severity of symptoms of allergic rhinoconjunctivitis during the ragweed blooming season. In the present study the complement-activating and specific IgE-binding capacities of various ragweed allergen preparations were compared. Elimination of physically adsorbed (flavonoid) pigments from the allergenic proteins had no significant effect on their complement-consuming capacity, although the process strongly diminished specific IgE binding. Removal of an IgE-binding trypsin inhibitor from RWA significantly enhanced RWA-induced complement activation, whereas it did not change IgE binding. These findings indicate that neither the physically adsorbed pigments nor the trypsin inhibitor are involved in complement activation by ragweed pollen allergens, and suggest that complement activation and specific IgE binding are distinct molecular properties of ragweed pollen allergen.
Corticosteroids restore the balance between locally produced Th1 and Th2 cytokines and immunoglobulin isotypes to normal in sarcoid lung
Authors: MILBURN H J , POULTER L W , DILMEC A , COCHRANE G M , KEMENY D M
Published: November 2003

In this study, we have investigated the balance between Th1- and Th2-like activity in the lungs in sarcoidosis and have determined the effect of corticosteroid treatment on this. Twenty-one patients with acute untreated sarcoidosis were investigated by bronchoalveolar lavage (BAL) and compared with 11 normal volunteers. Sixteen of the sarcoid patients required corticosteroid therapy and seven of these were reinvestigated after 2–3 months' treatment. In order to assess Th1- and Th2-like activity in the lungs, IgG subclasses and IgE were measured in BAL fluid and serum, and IL-2, IL-4 and interferon-gamma (IFN-γ) in BAL. In patients with untreated sarcoidosis, albumin-corrected BAL/serum ratios for IgG4 and IgE were significantly reduced (IgG4, 1·04±0·18 (mean±s.e.m.); IgE 9·58±3·11) compared with those in normal controls (IgG4 5·3±0·72, P<0·001; IgE 67·7±28·9, P<0·01). Estimates of actual levels of immunoglobulins produced in the lungs were also made and showed extremely high levels of total IgG in sarcoid patients (39·56±8·2 mg/l ) compared with controls (1·17±0·5 mg/l, P<0·001). Although there was no difference between the groups in amount of IgG4 locally produced, the proportion of total IgG which was IgG4 was greatly reduced in those with sarcoidosis (1·6±0·4% compared with 38·5±3·2%; P<0·001). Lavage levels of IL-4 were also reduced in sarcoid patients (IL-4 2·103±0·21 pg/ml) compared with those in normals (IL-4 6·8±1·05; P<0·001). Levels of IL-2 were lower (7·63±0·51 pg/ml compared with 9·4±0·95 pg/ml), but this difference was not significant. IFN-γ, however, could not be detected above 0·4 pg/ml in any of the normal lavage fluid, but was detectable in 12/21 patients with sarcoidosis (χ2=7·74; P<0·001). These changes reverted towards normal on treatment with oral corticosteroids. The mean albumin-corrected BAL/serum ratio for IgG4 before treatment was 0·88±0·33 compared with 5·5±2·1 (P<0·05) on treatment, and for IgE before treatment 9·52±2·15 compared with 50·8±17·9 (P<0·05) on treatment. Total IgG produced in the lung fell from 26·16±7·9 to 6·12±2·4 mg/l (P<0·001) on treatment, and the proportion of IgG4 locally produced rose from 2·3±0·8% to 23·9±6·1% (P<0·01). The mean level of IL-4 in lavage before treatment was 2·53±0·34 pg/ml compared with 4·7±0·34 (P<0·001) on treatment. Levels of IL-2 also rose significantly on treatment from 8·74±0·95 pg/ml before to 14·44±1·38 pg/ml (P<0·001) on treatment. Levels of IFN-γ fell from 1·65±0·43 pg/ml before treatment to undetectable levels in all patients (P<0·001) on treatment. These results demonstrate an imbalance between Th1- and Th2-like activity in the lungs in sarcoidosis, with suppression of Th2 and increase in Th1. Corticosteroid therapy restores the normal balance between Th1 and Th2 cytokines and immunoglobulins in the lungs, suggesting an effect on local immune regulation.
Detection of T cells reactive to intestinal alkaline phosphatase, an autoantigen in acute bacterial infections, and discrimination between autoantigen-specific CD5+ and CD5− B cells
Authors: KOLBUS N , FLEISCHER U , MÄDER M , FELGENHAUER K , BEUCHE W
Published: November 2003

Autoantibodies of the IgG isotype, specifically directed against intestinal alkaline phosphatase (IAP), occur transiently in the majority of sera from patients with acute bacterial infections. Sometimes they are observed in autoimmune diseases. Using a T cell proliferation assay, it was found that isolated peripheral blood mononuclear cells (PBMC) from IAP autoantibody (IAPA)-positive patients (n=18) responded significantly to IAP, whereas proliferation could not be induced in PBMC from healthy donors (n=11). Significant stimulation of PBMC from patients (n=11) was not obtained by use of transferrin, a common autoantigen in humans, indicating the specificity of stimulation of IAP-reactive T cells. Furthermore, T cell proliferation was observed when a highly purified IAP fragment (CNBr 21) spanning amino acids 334–462 of the primary structure of IAP was used as antigen. Thus, it was shown that an immunodominant T cell epitope resides within the CNBr 21 fragment which also contains a discontinuous B cell epitope as evaluated previously. Double immunocytochemical staining of T cell-depleted PBMC with IAP and an anti-human CD5 antibody allowed the detection of CD5+ B lymphocytes, which probably produce natural IAPA (nIAPA). These nIAPA-specific CD5+ B cells occurred with approximately the same frequency among T cell-depleted PBMC from healthy donors and those from patients. In contrast, IAPA-producing CD5− B cells were found in B cell-enriched preparations from patients, but not in those from healthy individuals.
Analysis of T cell activation pathways in patients with liver cirrhosis, impaired delayed hypersensitivity and other T cell-dependent functions
Authors: SCHIRREN C A , JUNG M-C , ZACHOVAL R , DIEPOLDER H , HOFFMANN R , RIETHMÜLLER G , PAPE G R
Published: November 2003

Patients with cirrhosis of the liver frequently demonstrate anergy in intracutaneous tests and fail to respond to vaccination, suggesting impaired delayed hypersensitivity and other T cell-dependent functions in vivo. T cell activation through the coordinated interaction of different cells of the immune system (B cell, antigen-presenting cells (APC)) is an important step in the induction of cellular and humoral immune responses. Impaired T cell-dependent functions in patients with liver cirrhosis may thus be explained by defective T cell activation. We prospectively investigated T cell activation pathways in 12 patients (nine males, three females) with alcoholic liver cirrhosis (seven Child Pugh stage A and B (CP A+B), five Child Pugh stage C (CP C)) and five healthy controls and compared the in vitro results of T cell activation with data obtained in vivo, e.g. intracutaneous tests and vaccination against hepatitis B surface antigen (HBs-Ag). Five out of eight patients who completed vaccination against hepatitis B virus infection were non-responders; one of the three responders had a non-protective anti-HBs titre. Moreover, three of five patients with alcoholic liver cirrhosis CP A+B, and two out of three with CP C were anergic in intracutaneous tests to a set of diverse antigens. All parameters of T cell activation were normal, including proliferation mediated by CD2, CD3–T cell receptor (TCR) complex, and CD28; acquisition of responsiveness to exogenous IL-2 and IL-4; activation of proteinkinase C (PKC) by phorbol ester and calcium influx by addition of ionomycin. The ability of monocytes to deliver costimulatory signals was preserved in patients with alcoholic cirrhosis. In addition, serum of patients with alcoholic liver disease did not inhibit T cell proliferation. We conclude that, although in patients with alcoholic liver cirrhosis T cell-dependent functions are impaired in vivo, T cell activation pathways are not responsible for the observed immune defect.
Chloroquine self-treatment and clinical outcome of cerebral malaria in children
Authors: PICOT S , NKWELLE AKEDE A , CHAULET J-F , TETANYE E , RINGWALD P , PRÉVOSTO J-M , BOUDIN C , AMBROISE-THOMAS P
Published: November 2003

Chloroquine is widely used as self-medication for presumptive treatment of malaria despite the existence of parasite resistance to the drug. Recent studies suggest that tumour necrosis factor-alpha (TNF-α) overproduction probably has a causal association with poor outcome in cerebral malaria. In addition, chloroquine has been shown to have inhibitory action on TNF-α synthesis. The present study aimed at evaluating chloroquine/TNF-α interaction in 90 children hospitalized for severe malaria in a malaria-endemic zone. TNF-α and chloroquine varied in the same range on admission, but there was an inverse correlation between the two: the higher the chloroquine level, the lower the TNF-α level. Parasite resistance to chloroquine in vitro was high. The clinical course in the patients was uneventful, save for two fatal cases and one survivor with neurological sequela. The above data suggest beneficial effects of chloroquine self-medication with respect to anti-TNF-α action. Rational use of this tool should be encouraged.
Long‐term follow‐up of patients with hyperprolactinaemia
Authors: Jeffcoate W. J. , Pound N. , Sturrock N. D. C. , Lambourne J.
Published: November 2003

AIM To determine the frequency with which hyperprolactinaemic illnes tends to resolve with time.

STUDY DESIGN A retrospective case‐notes review from a specialist endocrine unit in a provincial teaching hospital and tertiary referral centre.

PATIENTS Seventy women with hyperprolactinaemia referred to the unit in the 15 year period between May 1979 and May 1994. All those with a non‐pituitary cause or with macroadenoma had been excluded, as were those who did not have high‐resolution imaging, or who were on treatment at the time of referral.

INTERVENTION Intermittent course of treatment with dopamine receptor agonists according to individual need.

ENDPOINTS Latest serum PRL concentration in those who had discontinued treatment, and whether serum PRL tended to be lower in any particular group

RESULTS There was a significant fall in median PRL concentration from 2000 (714–8000) to 1000 mU/l (220–5600) in the 31 women who had discontinued therapy (P<0.0005), and serum PRL was normal (<700 mU/l) in 11 of them. Serum PRL also fell to normal in three of ten women who had no treatment at all. Final PRL concentration was normal in 35% of women who had had at least one pregnancy during the period of follow‐up compared to 14% who had not (P<0.05).

CONCLUSIONS These data confirm the findings of others that hyperprolactinaemia will prove self‐limiting in up to one‐third of women, and that pregnancy may be one factor which triggers a return to normal function
Androgens and the menopause; a study of 40–60‐year‐old women
Authors: Bancroft John , Cawood Elizabeth H. H.
Published: November 2003

OBJECTIVE The impact of the menopause on androgen production is poorly understood. We have investigated the impact of the menopause, as well as other factors such as age, body mass index (BMI) and cigarette smoking, on ovarian and adrenal androgen levels in women aged 40–60 years.

DESIGN Cross‐sectional study of blood hormones sampled weekly over one month in volunteer 40–60‐year‐old women.

SUBJECTS One hundred and forty‐one women, aged between 40 and 60, recruited from community sources (non‐clinical), not using hormone replacement or steroidal contraceptives, and with a current sexual partner. Fifty were categorized as premenopausal (ovulating), 37 as perimenopausal and 54 as post‐menopausal.

MEASUREMENTS The following variables were assessed; menopausal status (based on menstrual history and pattern and plasma progesterone), age, BMI, smoking, oestradiol (E2), oestrone (E1), LH, FSH, total testosterone (TT), androstenedione (A), SHBG, free androgen index (FAI), dihydroepiandrosterone (DHEA), dihydroepiandrosterone sulphate (DHEAS) and cortisol.

RESULTS are based on multiple regression analysis. TT was positively related to A, BMI and LH. A was negatively related to age and FSH, and positively to DHEA, DHEAS and premenopausal status. SHBG was negatively related to BMI and positively to E1 and non‐smoking. DHEA and DHEAS were negatively related to age and were higher in smokers. Both E1 and E2 were related to menopausal status and to FSH. Surprisingly, E2 was negatively related to BMI.

CONCLUSIONS A variety of factors influence androgen production in this age group. Whereas it is difficult to predict the effect of menopause on androgen levels, LH stimulation of post‐menopausal interstitial cells, modulated by a variety of factors including nutrition, and smoking, are likely to be relevant.
The assessment of growth hormone status in normal young adult males using a variety of provocative agents
Authors: Rahim A. , Toogood A. A. , Shalet S. M.
Published: November 2003

OBJECTIVE In adults, there are few data regarding GH responses to provocative stimuli other than insulin‐induced hypoglycaemia. We have compared the GH response to four different growth hormone secretagogues and placebo in normal healthy adult males.

DESIGN This was a prospective, randomized, placebo‐controlled study in 18 normal male subjects. After an overnight fast, an intravenous cannula was inserted into the arm of each subject and a blood sample was taken for GH at −30, −15, and 0 minutes. Four provocative agents (intravenous insulin 0.2 IU/kg; intravenous arginine 20 g/m2 as an infusion over 30 minutes; oral clonidine, either 100 or 200 μg; intramuscular glucagon 1 mg) and placebo were administered to each subject in a randomized manner on different days. Further blood samples were taken at 15‐minute intervals for 180 minutes for GH estimation.

RESULTS The median (range) GH peak response for each agent was insulin 107.7 (28.1–200) mU/l; arginine 22.3 (3.1–72.9) mU/l; glucagon 42 (11.8–200) mU/l; 100 μg clonidine 7.2 (<1–22.2) mU/l; 200 μg clonidine 8.2 (1.1–88) mU/l and placebo 2.4 (<1–30.2) mU/l. The peak GH response to insulin‐induced hypoglycaemia (ITT) was significantly greater than for any other agent (P<0.0001). The peak GH response to glucagon was significantly greater than for arginine (P<0.05), clonidine at 100 and 200 μg (P<0.01) and placebo (P<0.01). The peak GH response following administration of arginine was significantly greater than for clonidine 100 and 200 μg (P<0.05), and placebo (P<0.01). The peak GH response following clonidine 200 μg was not significantly greater than following clonidine 100 μg (P=0.38) or placebo.  On an individual basis two, six and 15 of 18 subjects failed to achieve a peak GH level of >20 mU/l to glucagon, arginine and clonidine respectively. In complete contrast only one subject achieved a peak response of less than 40 mU/l (28.1 mU/l) to ITT.

CONCLUSIONS The most profound GH release is seen after insulin‐induced hypoglycaemia. Glucagon appears to be more effective at inducing GH release than arginine. Clonidine at a dose of 100 or 200 μg is no more effective than placebo.
The short Synacthen and insulin stress tests in the assessment of the hypothalamic–pituitary–adrenal axis
Authors: Hurel S. J. , Thompson C. J. , Watson M. J. , Harris M. M. , Baylis P. H. , Kendall‐Taylor P.
Published: November 2003

OBJECTIVE The best dynamic test for the assessment of the hypothalamic–pituitary–adrenal axis and the interpretation of the cortisol levels, remain a matter of controversy. We aimed to establish normal ranges with current assays, for both the short Synacthen (SST) and insulin stress tests (IST) and then to use these data to examine whether the SST can satisfactorily substitute for the IST in assessment of the hypothalamic–pituitary–adrenal axis.

DESIGN Thirty SSTs and 27 ISTs were performed on different healthy volunteers. The results of all paired tests performed on patients in the last three years are reviewed.

SETTING Programmed Investigation Unit.

SUBJECTS Fifty‐seven healthy volunteers and 166 patients.

MAIN OUTCOME MEASURES Basal serum cortisol concentration and cortisol values obtained at 30 and 60 minutes during the SST compared to the maximum obtained with adequate hypoglycaemia (plasma glucose < 2mmol/l) during an IST.

RESULTS From normal data the mean‐2SD 30‐minute value during the SST was 392 nmol/l and 60‐minute value was 497 nmol/l. The maximal cortisol response (mean − 2SD) during the IST was 519 nmol/l. Sixty patients failed the IST, none of whom had a basal cortisol > 450 nmol/l and only six (10%) had a 30‐minute cortisol value > 600 nmol/l. The 30‐minute value provided a better index than the 60‐minute value. The basal, 30 and 60‐minute values during the SST all correlated positively and significantly with the maximal cortisol on IST. The correlations persisted for all microadenomas and macroadenomas secreting prolactin, gonadotrophins or growth hormone, patients undergoing either pre or post‐adenomectomy evaluation, and in those patients who had received long‐term steroids provided that the medication had been reduced and stopped two days prior to admission.

CONCLUSIONS Using a 30‐minute cortisol value > 600 nmol/l as a cut‐off, the short Synacthen test provides a suitable substitute for the insulin stress test. Adopting this policy will decrease the number of insulin stress tests performed by one‐quarter and thus provide a substantial saving without detriment to patient care.
Relation between serum interleukin‐6 and thyroid hormone concentrations in 270 hospital in‐patients with non‐thyroidal illness
Authors: Davies Peter H. , Black Elizabeth G. , Sheppard Michael C. , Franklyn Jayne A.
Published: November 2003

OBJECTIVES Non‐thyroidal illness (NTI) is frequently accompanied by alterations in circulating thyroid hormone concentrations, despite patients remaining clinically euthyroid. The mechanisms accounting for these changes in circulating thyroid hormone concentrations remain unknown. Much attention has focussed on the role of inflammatory cytokines which are known to be important mediators of disease. The aim of this study was to investigate the role of the cytokine interleukin‐6 (IL‐6) in alterations of thyroid hormone metabolism seen in NTI.

DESIGN Longitudinal study of hospital in‐patients, correlating serum IL‐6 concentrations with circulating thyroid hormone concentrations.

PATIENTS Two hundred and seventy in‐patients recruited consecutively, excluding those with known or suspected thyroid disorder. The patients were divided into 5 subgroups reflecting the nature of their NTI and comprised 41 patients with liver disease, 99 with renal disease, 19 intensive care (ITU) patients, 22 with cardiac disease and 89 patients with general medical, or surgical conditions.

MEASUREMENTS Serum IL‐6 concentrations were determined using a commercially obtained immunoassay (IL‐6 Quantikine assay, R&D Systems, Abingdon, UK). Serum total T4 and total T3 were measured using chemiluminescent immunometric assays (Kodak Clinical Diagnostics Ltd, Amersham, UK) and serum TSH was measured using a third‐generation chemiluminescent immunometric assay (Amerlite TSH 30, Kodak Clinical Diagnostics Ltd, Amersham, UK).

RESULTS Ninety‐three patients studied (35%) had a serum T3 below the normal range (<1.0 nmol/l), 89 patients (33%) had a serum T4 below the normal range (<65 nmol/l) and in 58 patients (21%) both serum T3 and T4 were below the normal range. There was a significant negative correlation between serum total T3 and IL‐6 (r = −0.219; P<0.001) and total T4 and IL‐6 (r = −0.132; P = 0.032), but not between TSH and IL‐6 (r = −0.075; P = 0.22). The ITU patient subgroup had the highest IL‐6 concentrations (229.3 ± 48.1 ng/l, mean ± standard error), whilst also having the lowest T3 (0.93 ± 0.08 nmol/l), TSH (0.79 ± 0.25 mU/l) and T4 concentrations (66.6 ± 7.3 nmol/l). The subgroup of patients under general medical or surgical care had least disturbance of their T3 (low in 19%) and T4 (low in 8%) concentrations, whilst also having the lowest mean IL‐6 concentration (39.0 ± 5.3 ng/l). The renal patient subgroup, whilst including a high proportion of patients with low T3 (39%) and T4 (45%) concentration, demonstrated only modest elevation of IL‐6 concentrations (mean 41.4 ± 8.5 ng/l).

CONCLUSIONS Our data revealed a statistical relation between elevated serum IL‐6 concentrations and alterations in circulating thyroid hormone concentrations seen in NTI; however, the findings in patients with renal disease suggest that circulating IL‐6 is not the only factor responsible for alteration in thyroid hormone metabolism in NTI.
Pre‐clinical Cushing's syndrome: an unexpected frequent cause of poor glycaemic control in obese diabetic patients
Authors: Leibowitz Gil , Tsur Anat , Chayen Susan D. , Salameh Mohammad , Raz Itamar , Cerasi Erol , Gross David J.
Published: November 2003

OBJECTIVE Autonomous cortisol secretion without clinical stigmata of Cushing's syndrome (CS) has been recently recognized and termed pre‐clinical or sub‐clinical CS. The common assumption is that CS is an extremely rare cause of uncontrolled diabetes; however, the prevalence of this entity has not been studied. We assessed the prevalence of pre‐clinical CS among obese patients with uncontrolled diabetes.

PATIENTS AND DESIGN (1) In a retrospective analysis, the medical records of 63 patients with endogenous CS were reviewed. (2) In a cross‐sectional study, 90 obese patients (BMI >25 kg/m2) followed in a University Hospital and the local Health Fund endocrine and diabetes clinics, with poorly controlled diabetes (glycosylated haemoglobin >9%), underwent an overnight 1 mg dexamethasone suppression. In patients with non‐suppressible cortisol levels (>140 nmol/l), Liddle's 2 and 8 mg dexamethasone suppression tests and imaging studies were performed.

MEASUREMENTS The prevalence of poorly controlled diabetes, the major presenting symptom of CS, was assessed in the retrospective analysis. The prevalence of ‘true’ CS and the false positive rate in the overnight dexamethasone suppression test were calculated. The endocrine evaluation of the patients with pre‐clinical CS and the effects of surgical cure on glycaemic control are described.

RESULTS In the retrospective analysis, 11 (17.5%) had diabetes and 2 (3.2%) lacked the classic physical characteristics of the syndrome. In the cross‐sectional study, 4 patients failed to suppress plasma cortisol (<140 nmol/l). In one patient the diagnosis of CS was not confirmed by a standard Liddle’s test and was therefore considered false positive. In the other 3, the diagnosis of CS was confirmed (prevalence of 3.3%, 95% confidence interval 1–9%). In all other patients the overnight cortisol suppression test was normal (cortisol level 47.3 ± 2.5 nmol/l (mean ± SEM)). After surgical treatment of CS, glycaemic control was markedly improved in all 5 patients (2 from retrospective and 3 from cross‐sectional studies).

CONCLUSIONS The prevalence of pre‐clinical Cushing's syndrome in obese patients with poorly controlled diabetes appears to be considerably higher than previously believed. The overnight dexamethasone suppression test proved to be a simple, sensitive and highly specific screening test for Cushing's syndrome despite the presence of obesity and hyperglycaemia.
The outcome of hypophysectomy for prolactinomas in the era of dopamine agonist therapy
Authors: Soule Steven G. , Farhi Jacob , Conway Gerard S. , Jacobs Howard S. , Powell Michael
Published: November 2003

OBJECTIVE Dopamine agonists are the primary therapeutic modality for the majority of patients with prolactinomas, with pituitary surgery reserved for those patients intolerant of or resistant to these agents. Most published surgical series, however, contain patients treated by surgery as the primary therapeutic modality. Previous exposure to dopamine agonists or the selection of patients with prolactinomas resistant to conventional therapy may potentially compromise the surgical success rate. The purpose of this study was to evaluate the efficacy and safety of pituitary surgery for prolactinomas in a tertiary referral centre where the majority of patients were operated on after treatment with dopamine agonists.

DESIGN A retrospective review of the outcome of pituitary surgery for prolactinomas performed at a tertiary neurosurgical centre by a single neurosurgeon.

PATIENTS Twenty‐three patients underwent excision of a macro and 11 excision of a micro‐prolactinoma.

MEASUREMENTS Pituitary tumour diameter was determined by CT or MRI imaging. Pre and post‐operative measurements were made of serum PRL concentration (off dopamine agonist therapy), free T4, free T3, LH and testosterone (males). Post‐operative restoration of a menstrual cycle was taken to indicate resolution of hypogonadism in female patients.

RESULTS The majority (73.9%) of the patients with macro and all with micro‐prolactinomas had received dopamine agonists preoperatively. Of the 23 patients with macroprolactinomas, in whom the median pre‐operative PRL concentration was 13255 mU/l, 17 (73.9%) had radiological evidence of suprasellar extension and 5 (21.7%) cavernous sinus invasion. Only 4 (17.4%) of the patients with macroprolactinomas had a normal serum PRL post‐operatively, although there was an improvement in visual fields in 66% of those with preoperative defects. The median preoperative PRL concentration was 4309 mU/l in the patients with microprolactinomas, significantly lower than in the macroprolactinoma group (P = 0.02). Despite a significant fall in serum PRL post‐operatively (median PRL 860 mU/l, P = 0.0001), only 45.5% of patients had a normal serum PRL concentration after surgery.

CONCLUSIONS The cure rate following pituitary surgery for prolactinomas in a tertiary referral centre was low when compared with previous series in which surgery was used as the primary therapeutic modality. We suggest this may result both from dopamine agonist pretreatment and the referral of prolactinomas resistant to conventional therapy. The outcome is probably a more realistic reflection of the results of pituitary surgery for prolactinomas as currently practised in the majority of neuroendocrine centres.
Glucose tolerance, insulin secretion, insulin sensitivity and glucose effectiveness in normal and overweight hyperthyroid women
Authors: Gonzalo Ma. Angeles , Grant Carlos , Moreno Inmaculada , Garcia Fernando J. , Suárez Ana Isabel , Herrera‐Pombo Jose Luis , Rovira Adela
Published: November 2003

OBJECTIVE Inter‐relationships between insulin sensi‐tivity and body weight in patients with hyperthyroidism remain incompletely understood. We have examined whether a mild excess of body weight exacerbates the metabolic abnormalities of spontaneous hyperthyroidism.

DESIGN AND PATIENTS Insulin‐modified intravenous glucose tolerance tests were performed on 14 hyperthy‐roid women with body mass indices (BMI) ranging from 21 to 31 kg/m2. A control group of 19 healthy women matched for age and BMI was also studied.

MEASUREMENTS Intravenous glucose tolerance (KG), first and second‐phase integrated insulin responses to glucose, the integrated glucose area under the curve (AUC), and minimal model parameters of insulin sensitivity (SI) and glucose effectiveness (SG) were determined.

RESULTS Hyperthyroid women had mean KG, glucose‐induced insulin secretion and SG values similar to those in control women. The mean glucose AUC was higher in hyperthyroid patients (P<0.05). Lower insulin sensitivity was observed in hyperthyroid patients than in control women (SI=0.38±0.07 vs 0.59±0.07 l/min pmol 104 (mean±SEM), P<0.05). A steeper decline in insulin sensitivity with increase in body mass index was found in hyperthyroid women when compared with the control group, after adjusting for age. When groups were compared according to their BMI, hyperthyroid women with normal weight (BMI≤25 kg/m2, n=8) had mean KG, insulin response to glucose, glucose AUC, SG and SI values similar to those in normal weight control women (n=11). Overweight hyperthyroid patients (BMI>25 kg/m2, n=6) had a higher (P<0.05) second‐phase insulin response to glucose than normal weight patients, a higher glucose AUC (P<0.05) than normal weight patients and overweight controls (n=8), and a lower SI (P<0.05) than normal weight patients and overweight controls. SG was not influenced by BMI in hyperthyroid patients.

CONCLUSIONS These results suggest that overall glucose tolerance was not significantly affected in normal weight hyperthyroid women. However, when a moderate excess of weight is also present, a state of clear insulin resistance occurs.
Circulating inhibins and activin A during GnRH‐analogue down‐regulation and ovarian hyperstimulation with recombinant FSH for in‐vitro fertilization‐embryo transfer
Authors: Lockwood G. M. , Muttukrishna S. , Groome N. P. , Knight P. G. , Ledger W. L.
Published: November 2003

OBJECTIVE We have investigated serial changes in plasma concentrations of inhibin A, inhibin B, pro αC and activin A in women undergoing stimulation with recombinant FSH in ‘long‐protocol’ down‐regulated cycles of IVF treatment.

DESIGN Blood samples were collected during the entire IVF treatment cycle at points coinciding with the early follicular phase of the cycle preceding treatment, pituitary down‐regulation, stimulation with recombinant FSH, ovulatory triggering, and the luteal phase of the cycle. In patients who achieved conception, blood samples were also taken during the first 2 weeks of pregnancy. All samples were analysed for inhibin A, inhibin B, pro αC, activin A and oestradiol.

PATIENTS Fifteen women with normal ovarian function undergoing IVF treatment with tubal factor, mild endometriosis or idiopathic infertility.

RESULTS During pituitary desensitization, both inhibin A and inhibin B were significantly (P<0.001, P=0.002, respectively) reduced whereas levels of pro αC and activin A were largely unaltered. Levels of both inhibins rose markedly (P<0.01) during FSH stimulation and a further rise in inhibin A was detected on the day after ovulatory trigger. Levels of both inhibin A and inhibin B then fell during and after oocyte pickup and continued to fall during the luteal phase. Activin A levels rose less markedly during gonadotrophin stimulation. Statistical analysis showed a high degree of correlation between the number of follicles (>10 mm) and serum inhibin A (r=0.65, P<0.01) and pro αC (r=0.65, P<0.01) concentrations during the late follicular phase.

CONCLUSIONS These results indicate that ovarian production of dimeric inhibin A and B are gonadotrophin dependent, whereas activin A may have a significant gonadotrophin independent or extra‐gonadal source. Inhibin A and pro αC may be useful markers for monitoring the effects of gonadotrophin stimulation.
Size at birth and adrenocortical function in childhood
Authors: Clark Phillipa M. , Hindmarsh Peter C. , Shiell Alistair W. , Law Catherine M. , Honour John W. , Barker David J. P.
Published: November 2003

OBJECTIVE The mechanisms underlying the association between reduced size at birth and cardiovascular disease and non‐insulin‐dependent diabetes mellitus in adult life are not known. One possibility is that the intra‐uterine environment has permanent effects on the function or activity of the hypothalamo‐pituitary‐adrenal axis. We tested this by relating size at birth to the urinary excretion of adrenal androgen and glucocorticoid metabolites in a population sample of 9‐year‐old children.

SUBJECTS AND METHODS One hundred and ninety children (89 boys and 101 girls) of known present height, weight and size at birth collected a 24‐hour urine sample. The urinary breakdown products of dehydroepiandrosterone sulphate and of cortisol and cortisone were measured by gas chromatography and their respective breakdown products summed (‘adrenal androgen metabolites’ and ‘glucocorticoid metabolites’). Excretion was expressed in μg/day.

RESULTS Urinary adrenal androgen metabolite excretion was higher in children who had been light at birth. A 1‐kg decrease in birthweight was associated with a 40% (95% CI 9–79%) increase in metabolite excretion. Excretion was positively associated with current weight and age, but the relation with birth weight was independent of weight, age or sex. Urinary glucocorticoid metabolite excretion was positively associated with current weight, but not independently with age. The urinary excretion of total glucocorticoid metabolites was higher in children who had been light at birth, but the relation was best described as U‐shaped, with the highest average urinary glucocorticoid metabolite excretion being found in children who had been either light or heavy at birth. The U‐shaped (quadratic) relation persisted after adjustment for sex and current weight (P for quadratic term 0.006).

CONCLUSION These findings suggests that the intra‐uterine environment, as measured by fetal size at birth, has long‐lasting effects on the function of the hypothalamo‐pituitary‐adrenal axis.
Cushing's syndrome secondary to ectopic ACTH secretion from a primary ovarian carcinoma
Authors: Ball S. G. , Davison J. M. , Burt A. D. , McNicol A. M. , Baylis P. H.
Published: November 2003

We report a 63‐year‐old woman who presented with clinical and biochemical features of ACTH dependent Cushing's syndrome secondary to a primary ovarian carcinoma. The tumour produced very high levels of ACTH precursors, consistent with defective POMC processing.