bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential graft-versus-leukaemia effect by CD28 and CD40 co-stimulatory blockade after graft-versus-host disease prophylaxis
Authors: OHATA J , SAKURAI J , SAITO K , TANI K , ASANO S , AZUMA M
Published: March 2003

Co-stimulatory blockade may be a promising strategy for tolerance induction in transplantation. In allogeneic bone marrow transplantation (BMT) for leukaemia treatment, however, preservation of the graft-versus-leukaemia (GVL) effect is another critical requirement for clinical application. In this study, we have compared the effect on GVL of using CD28 and CD40 co-stimulatory blockades as graft-versus-host disease (GVHD) prophylaxis in a murine allogeneic BMT model with simultaneous transfer of BCL1 leukaemia. Despite the relative improvement of GVHD as assessed by survival and body weight in both treatment regimes, treatment with anti-CD154 moAb clearly diminished the GVL effect, whereas treatment with anti-CD80 and CD86 MoAbs maintained this effect. Although T cell-mediated effector function at 14 days post-BMT assessed by IFNγ expression and cytotoxicity against host alloantigen was comparable between both co-stimulatory blockades, IL-12 mRNA expression was preferentially reduced by CD40 blockade. Our results suggest the differential involvement of the CD28 and CD40 co-stimulatory pathways in the development of GVHD and GVL effects. CD28 blockade may be a favourable strategy for tolerance induction in leukaemia patients undergoing BMT.
The molecular mechanism in activation-induced cell death of an Ag-reactive B cell clon
Authors: HAMANO T , IWASAKI T , OGATA A , HASHIMOTO N , KAKISHITA E
Published: March 2003

TPA-1 is a subclone of B cell hybridomas established by somatic hybridization using B cells of A/J mice immunized with TNP-LPS, and expresses a receptor for TNP on the cell membrane. The present study showed that TPA-1 was induced to apoptotic cell death upon treatment with TNP-BSA. Therefore, TPA-1 is considered to provide a good model for the study on activation-induced cell death of mature B cells induced by soluble antigen. TNP-BSA treatment caused the generation of a large amount of intracellular reactive oxygen species (ROS) of TPA-1, and the addition of the monovalent thiol-reactive compound: monochlorobimane (MCB) rescued it from apoptosis as well as the antioxidant reagent: N-acetyl-l-cysteine. Furthermore, MCB markedly inhibited the generation of ROS and prevented the disruption of mitochondrial membrane potential that was induced by TNP-BSA treatment. In addition, it counteracted the effect of TNP-BSA on the expression of the Bcl-2 family, resulting in down-regulation of Bax and Bad and up-regulation of Bcl-XL. Taken together, these results suggest strongly that oxidative stress of mitochondria may be involved directly in apoptotic cell death by engagement of antigen receptors on mature B cells with soluble antigen.
Impaired up-regulation of CD70 and CD86 in naive (CD27−) B cells from patients with common variable immunodeficiency (CVID)
Authors: GROTH C , DRÄGER R , WARNATZ K , WOLFF-VORBECK G , SCHMIDT S , EIBEL H , SCHLESIER M , PETER H-H
Published: March 2003

CVID is characterized by reduced serum levels of all switched immunoglobulin isotypes (IgG, IgA, IgE) predisposing patients to recurrent infections of their respiratory and gastrointestinal tract. Correspondingly, most CVID patients exhibit a severely decreased proportion of class switched memory B cells (CD19+CD27+IgD−IgM−IgG+ or IgA+) in their peripheral blood (CVID type I). We previously identified a subgroup of CVID patients showing a significantly reduced expression of CD86 and CD137 following activation in vitro of PBMC or purified B cells (CD19+) with anti-IgM plus IL-2. Here we extend our previous studies by asking whether highly purified, cell-sorted naive B cells show already an expression defect of B cell surface molecules relevant in activation (CD39, CD69), differentiation (CD24, CD27, CD38) or T–B interaction (CD25, CD70, CD86). We stimulated cell-sorted, naive B cells (CD19+CD27−IgM+IgDhighIgG−IgA−) from 10 CVID patients and 10 healthy controls for 4 days with anti-IgM plus IL-2 in the absence or presence of autologous CD4+ T cells and measured the expression of the referred surface molecules. Based on reduced or normal numbers of switched memory B cells the CVID patients had previously been classified into eight type I patients and two type II patients, respectively. Interestingly, only the molecules CD25, CD70 and CD86, all relevant in cognate T–B interaction, showed a significantly lower expression in naive B cells from CVID patients compared to controls. While coculture with autologous CD4+ T cells normalized the CD25 expression, CD70 and CD86 expression remained subnormal, notably in the eight CVID patients of type I. These findings strongly suggest an intrinsic signalling or expression defect for CD70/CD86 at the level of naive B cells in type I CVID patients.
Systemic treatment with anti-CD40 antibody stimulates Langerhans cell migration from the skin
Authors: JOLLES S , CHRISTENSEN J , HOLMAN M , KLAUS G B , AGER A
Published: March 2003

Epidermal Langerhans cells (LCs) play a pivotal role in the initiation of cutaneous immune responses. The maturation of LCs and their migration from the skin to the T cell areas of draining lymph nodes are essential for the delivery and presentation of antigen to naïve T cells. CD40, which acts as a costimulatory molecule, is present on LCs and the basal layer of keratinocytes in the skin. We show here that systemic treatment of mice with anti-CD40 antibody stimulates the migration of LCs out of the epidermis with a 70% reduction in LC numbers after 7 days, although changes in LC morphology are detectable as early as day 3. LC numbers in the epidermis returned to 90% of normal by day 21. As well as morphological changes, LC showed up-regulated levels of Class II and ICAM-1, with only minimal changes in CD86 expression 3 days following anti-CD40 treatment. Despite increased levels of Class II and ICAM-1, epidermal LC isolated from anti-CD40 treated mice were poor stimulators of a unidirectional allogeneic mixed leucocyte reaction (MLR), as were epidermal LC isolated from control mice. These results indicate that CD40 stimulation is an effective signal for LC migration, distinct from maturation of immunostimulatory function in the epidermis, which is not altered. These observations may have important implications for the mechanism of action of agonistic anti-CD40 antibodies, which have been used as an adjuvant in models of infection and experimental tumours and the primary immunodeficiency Hyper IgM syndrome caused by deficiency of CD40 ligand.
T-cells in the cerebrospinal fluid express a similar repertoire of inflammatory chemokine receptors in the absence or presence of CNS inflammation: implications for CNS trafficking
Authors: KIVISÄKK P , TREBST C , LIU Z , TUCKY B H , SØRENSEN T L , RUDICK R A , MACK M , RANSOHOFF R M
Published: March 2003

It is believed that chemokines and their receptors are involved in trafficking of T-cells to the central nervous system (CNS). The aim of the current study was to define the expression on cerebrospinal fluid (CSF) T-cells of six chemokine receptors associated with trafficking to sites of inflammation. Flow cytometry was used to detect chemokine receptor expression. We observed that CD3+T-cells in the CSF express a restricted array of inflammatory chemokine receptors, specifically CXCR3, CCR5 and CCR6, but little CCR1-3. This repertoire was independent of the presence of CNS inflammation, since comparable findings were obtained in patients with multiple sclerosis (MS) and individuals with non-inflammatory neurological diseases. The enrichment of CCR5+T-cells in the CSF could largely be explained by higher frequency of CD4+/CD45RO+T-cells in this compartment. In contrast, CD4+/CD45RO+T-cells expressing CXCR3 were significantly enriched in CSF as compared with blood. Similar levels of CCR6+/CD3+T-cells were observed in blood and CSF, while levels of CCR2+/CD3+T-cells were lower in CSF than in blood. The CSF was virtually devoid of CCR5+/CXCR3- T-cells, suggesting that the expression of CCR5 alone is not sufficient for the trafficking of CD3+T-cells to the CSF. We hypothesize that CXCR3 is the principal inflammatory chemokine receptor involved in intrathecal accumulation of T-cells in MS. Through interactions with its ligands, CXCR3 is proposed to mediate retention of T-cells in the inflamed CNS.
The immunology of primary biliary cirrhosis: the end of the beginning?
Authors: PALMER J M , KIRBY J A , JONES D E J
Published: March 2003

The chronic liver disease primary biliary cirrhosis (PBC) is characterised by autoreactive B-cell and T-cell responses directed against mitochondrial antigens. In recent years these responses have been extensively characterised and the principal PBC associated autoantigen identified as pyruvate dehydrogenase complex (PDC). The identification of anti-PDC responses (present in over 95% of PDC patients) has given rise to important questions pertinent to our understanding of the pathogenesis of PBC. What specific role to anti-PDC responses play in target cell damage? How and why does immune tolerance break down to as highly conserved and ubiquitously expressed self-antigen as PDC? Why does breakdown in tolerance to an antigen present in all nucleated cells result in damage restricted to the intra-hepatic bile ducts? In attempting to answer these key questions we have, in this review, proposed a unifying hypothesis for the pathogenesis of PBC.
Ribavirin increases mitogen- and antigen-induced expression of CD40L on CD4+ T cellsin vivo
Authors: BERGAMINI A , CEPPARULO M , BOLACCHI F , ARACO A , TISONE G , OMBRES D , ROCCHI G , ANGELICO M
Published: March 2003

Here, CD40L expression and cytokine production have been analysed in peripheral blood cells from orthotopic liver transplantation (OLT) recipients treated with ribavirin for recurrent chronic hepatitis C. The study included 18 OLT recipients treated with ribavirin, eight control OLT recipients and 10 healthy controls. FACS analysis showed that baseline expression of CD40L was not different between ribavirin-treated patients and controls. In contrast, after stimulation with both HCV core antigen and phorbol myristate acetate (PMA) plus ionomycin (IO), the expression of CD40L on CD4 lymphocytes was significantly higher in the ribavirin group compared with controls. In the ribavirin group, the increased expression of CD40L significantly correlated with reduction of HCV RNA levels with respect to pretreatment values. Finally, ribavirin treatment was not associated with modification of PMA-IO-induced cytokine production by T lymphocytes and interleukin (IL)-1β and tumour necrosis-α (TNF)-α production by CD40L-stimulated monocytes. In conclusion, these data indicate that ribavirin ­upmodulates CD40L expression on CD4 T cells, a property which may account in part for its ability to enhance the antiviral activity of interferon-α in the treatment of chronic HCV infection.
Impact of NNRTI compared to PI-based highly active antiretroviral therapy on CCR5 receptor expression, β  -chemokines and IL-16 secretion in HIV-1 infection
Authors: BURTON C T , HARDY G A D , SULLIVAN A K , NELSON M R , GAZZARD B , GOTCH F M , IMAMI N
Published: March 2003

Interleukin-16 (IL-16) and the β-chemokines (RANTES, monocyte chemotactic protein-1 (MCP-1), macrophage inhibitory protein (MIP)-1α and (MIP)-1β) are soluble in vitro suppressors of macrophage tropic HIV-1 strains. The reduction of HIV-1 RNA plasma levels in late-stage patients receiving protease inhibitors has been associated with increased concentrations of MIP-1α, MIP-1β, RANTES and IL-16 and a decrease in levels of MCP-1. We determined plasma levels of MCP-1, MIP-1α, MIP-1β, RANTES and IL-16 during the first 16 weeks of highly active antiretroviral therapy (HAART) in chronic HIV-1-infected patients. Patients were administered one of two therapeutic regimens based on either   a   protease   inhibitor (PI) or   a   non-nucleoside   reverse   transcriptase    inhibitor (NNRTI).   No differences were seen in the levels of RANTES and IL-16 over the first 16 weeks of HAART in either treatment group. MCP-1 decreased significantly in the PI-treated group over the first 16 weeks of HAART (P = 0·0003). A significant increase was observed in the levels of MIP-1α and MIP-1β in the NNRTI cohort (P = 0·0010 and P = 0·0012, respectively). A significant decrease in levels of MIP-1α and MIP-1β (P = 0·0015 and P = 0·0299, respectively) was observed over the 16 weeks in the PI cohort. A significant difference was seen when the levels of MIP-1α and MIP-1β were compared between the NNRTI and the PI cohorts at week 16 (P = 0·04 and P = 0·05, respectively). Evaluation of CCR5 expression ex vivo revealed no difference between the two treatment groups. Patients were genotyped for CCR5 Δ32 and the incidence of heterozygosity was lower than in the HIV-1 seronegative controls (3% compared to 19%).
Detection and characterization of plasma cells in peripheral blood: correlation of IgE+ plasma cell frequency with IgE serum titre
Authors: HORST A , HUNZELMANN N , ARCE S , HERBER M , MANZ R A , RADBRUCH A , NISCHT R , SCHMITZ J , ASSENMACHER M
Published: March 2003

In atopic patients and patients with hyper-IgE syndrome (HIE) highly elevated IgE serum levels can be detected. Due to their very low frequency little is known about IgE-producing plasma cells (PC) in peripheral blood. We used CD138 MACS microbeads to enrich plasma cells from peripheral blood of normal donors, atopic patients and one HIE patient. CD138+ cells were mainly CD45+, CD44++, CD19dim, CD38++, CD27++, CD86+, HLA-DR+/++, CD71dim, VLA-4+, VLA-5–, CD28–, CD25–, CD69–, CLA–, CD20–, CD21– and CD22–. They show weak expression of surface Ig but high levels of intracellular Ig and they secrete Ig in culture. Thus CD138+ cells from peripheral blood show characteristics of early plasma cells. IgE+ CD138+ plasma cells could be detected in 19 of 24 normal donors with an average frequency of 0·06% IgE+ cells among CD138+ cells. Higher frequencies were detected in atopic patients, atopic patients with markedly elevated serum IgE levels and the hyper-IgE patient with an average of 0·32%, 7·21% and 6·54%, respectively. Additionally, using the recently developed cellular affinity matrix technology, we were able to detect IgE secreting plasma cells and thereby could demonstrate that most of the IgE secreting cells express CD138. The frequency of IgE+ CD138+ cells among PBMC correlated highly significantly with serum IgE titres (r = 0·8532***), indicating that IgE secreting CD138+ cells in peripheral blood are directly related to the plasma cell pool contributing to the IgE titre.
Monocytes are primed to produce the Th1 type cytokine IL-12 in normal human pregnancy: an intracellular flow cytometric analysis of peripheral blood mononuclear cells
Authors: SACKS G P , REDMAN C W G , SARGENT I L
Published: March 2003

This paper considers both monocytes and peripheral blood lymphocytes as potential targets for maternal immunological modulation in pregnancy. Peripheral blood mononuclear cells (PBMCs) from non-pregnant and normal pregnant donors were stimulated in vitro, and cytokine production detected intracellularly by flow cytometry. It was found that monocyte production of TNF-α was unaltered in pregnancy, while production of IL-12 was significantly enhanced. In contrast, production of the Th1 type cytokine IFN-γ was suppressed in the lymphocyte subsets: CD4+ T helper cells and CD56+ NK cells. Production of the Th2 type cytokine IL-4 in CD4+ cells was not significantly altered in pregnancy. These data suggest that the concept that pregnancy is a ‘Th2 phenomenon’ cannot be generalized to the function of all aspects of maternal cellular immunity as, paradoxically, circulating monocytes are ‘primed’ to produce the Th1 cytokine IL-12. Furthermore, these data support the hypothesis that components of maternal innate immunity are activated in normal pregnancy.
A very high dose dexamethasone suppression test for differential diagnosis of Cushing's syndrome
Authors: Al‐Saadi  , Diederich  , Oelkers
Published: March 2003

OBJECTIVE

The high‐dose dexamethasone (dex) suppression test of cortisol secretion (8×2 mg dex over two days or 8 mg overnight) is a mainstay in the differential diagnosis of Cushing's syndrome (CS). In some patients with pituitary Cushing's disease (CD), however, plasma cortisol is not suppressed to <50% of control by 8 mg of dex. We therefore hypothesized that a higher dose of dex might produce more effective suppression of cortisol secretion in CD.

DESIGN AND SUBJECTS

We routinely tested the diagnostic efficacy of a very high dose of dex (32 mg, i.e. 4×8 mg in 24 hours) in comparison with the 8 mg overnight dex test in a population of patients with CD, in which an unusually high percentage was refractory to 8 mg dex. End points were the suppression of plasma cortisol, plasma ACTH and urinary free cortisol (UFC) to <50% of control. Corticotrophin releasing hormone (human CRH) tests were also performed.

RESULTS

Eleven out of 26 (11/26) patients with CD (42%), among them six with pituitary macroadenomas, failed to show suppression of plasma cortisol after 8 mg dex. Five out of 19 patients (26%) with CD failed to suppress after 32 mg dex. Only 3/19 (16%) failed to suppress UFC after 32 mg dex. In non‐ pituitary CS (n=11), only one patient with macronodular adrenal hyperplasia showed significant suppression of plasma cortisol, but not UFC, after 32 mg dex. ACTH suppression after 8 or 32 mg dex was often less pronounced than that of cortisol and was of no diagnostic value. Cortisol stimulation by 23% after hCRH injection differentiated 100% of patients with CD from other forms of CS.

CONCLUSION

In this series, the hCRH test was the most reliable test for the differential diagnosis of Cushing's syndrome. The 32 mg dexamethasone test with measurement of urinary free cortisol was clearly superior to the 8 mg test and to other aspects of the very high dose dexamethasone test. It can be recommended for ‘non‐suppressible’ patients with ACTH‐dependent Cushing's syndrome and can be performed on outpatients.
Enhanced growth hormone (GH) responsiveness to GH‐releasing hormone after dietary restriction in patients with Cushing's syndrome
Authors: Leal‐Cerro  , Venegas  , Garcia‐Pesquera  , Jimenez  , Astorga  , Casanueva  , Dieguez
Published: March 2003

OBJECTIVE

In patients with Cushing's syndrome, decreased growth hormone (GH) secretion is observed although the basic mechanism is not yet understood. A short‐term hypocaloric diet is known to increase both spontaneous and GHRH‐stimulated GH secretion in normal subjects. In order to gain further insight into the altered GH secretion in patients with Cushing's syndrome, we assessed the effect of a short‐term hypocaloric diet on GH responses to GHRH in these patients.

DESIGN

Two GHRH tests (1 μg/kg i.v.) were performed, the first under basal conditions (normocaloric diet) and the second after a 3‐day hypocaloric diet (650 cal/day).

PATIENTS

Six female patients with untreated Cushing's disease.

MEASUREMENTS

Plasma GH levels were measured by immunoradiometric assay.

RESULTS

GHRH‐induced GH release was impaired in patients with Cushing's disease on a normal diet (mean peak 12.4 ± 6.4 mU/l, area under the curve (AUC) 744 ± 332 mU/l/120 min). Following a hypocaloric diet, GH responses to GHRH were markedly enhanced in the same group of patients (mean peak 46.2 ± 14.8 mU/l, AUC 3142 ± 1032 mU/l/120 min, P < 0.05).

CONCLUSIONS

This study demonstrates that in patients with Cushing's disease the somatotroph hyporesponsiveness to growth hormone releasing home is improved after a short‐term hypocaloric diet. Therefore, blunted growth‐hormone secretion in chronic hypercortisolism is a potentially reversible state and the secretory capacity of the somatotroph appears not to be severely compromised in patients with Cushing's disease.
The tender neck: thyroiditis or thyroid abscess?
Authors: Houghton David J. , Gray Henry W. , MacKenzie Kenneth
Published: March 2003

Following apparent subacute thyroiditis, a 16‐year‐old girl developed a left thyroid abscess thought to be secondary to steroids and haematogenous spread from a pilonidal abscess. The thyroid suppuration became recurrent and required partial thyroidectomy. Further left‐sided abscess formation in the neck prompted a barium swallow which revealed the source of infection to be a sinus tract arising from the left piriform fossa. The patent fourth branchial sinus tract was later excised. All patients with a tender thyroid should have ultrasound‐guided fine needle aspiration to establish the diagnosis. If suppuration is confirmed, a barium swallow is advised to exclude a sinus tract from the piriform fossa.
Exaggerated 17‐hydroxyprogesterone response to short‐term adrenal stimulation and evidence for CYP21B gene point mutations in true precocious puberty
Authors: Cisternino Mariangela , Dondi Elisabetta , Martinetti Miryam , Lorini Renata , Salvaneschi Laura , Cuccia Mariaclara , Severi Francesca
Published: March 2003

OBJECTIVE

Following the observation of two patients affected by true precocious puberty who went on to develop polycystic ovary syndrome (PCOS) and who were found to be heterozygotes (carriers) for 21‐hydroxylase deficiency (21‐OHD), we decided to evaluate the frequency of heterozygosity for adrenal 21‐OHD in patients with true precocious puberty.

STUDY DESIGN

We investigated 32 girls affected by true precocious puberty, by the single‐dose ACTH stimulation test, HLA typing and the molecular analysis of the CYP21B gene encoding for the 21‐OH enzyme, in order to detect gene deletions or point mutations. Twenty‐eight cases were on LHRH analogue treatment and the remaining four, untreated owing to parental refusal, were investigated 0.5–1.5 years after spontaneous menarche.

RESULTS

After ACTH testing, 13 out of the 32 (41%) cases displayed higher 17‐hydroxyprogesterone (17‐OHP) levels than normal but less than those found in patients affected by nonclassical adrenal hyperplasia (CAH); these levels were similar to those observed in obligate heterozygotes for CAH due to 21‐hydroxylase deficiency (21‐OHD). HLA typing showed a significantly increased frequency of the HLA alleles A28 and B14 which are peculiar to the HLA haplotypes of nonclassical CAH due to 21‐OHD. Molecular analysis of the CYP21B gene showed that in four out of the 10 tested patients with an exaggerated 17‐OHP response there were heterozygous point mutations of the CYP21B gene. In contrast, no CYP21B gene abnormalities were detected in the eight tested patients with normal 17‐OHP. No differences were found between carriers and non‐carriers of the 21‐OHD with regard to age at onset of precocious puberty, clinical features, bone age acceleration and gonadal suppression induced by LH‐RH analogue treatment. Two out of the four untreated patients who were investigated after menarche were found to be carriers of the 21‐OHD; these girls showed signs of androgen excess, irregular menses and polycystic ovaries.

CONCLUSIONS

A high frequency of heterozygosity for adrenal steroid 21‐OHD was found in our patients with true precocious puberty. This adrenal defect does not seem to influence the pattern of central precocious puberty, but these patients require long‐term follow‐up as they might go on to develop polycystic ovary syndrome (PCOS). Whether or not heterozygosity of the 21‐OHD may be related to the premature activation of the hypothalamo‐pituitary‐gonodal axis remains to be defined.
Giant cell granulamatous hypophysitis with remarkable uptake on Gallium‐67 scintigraphy
Authors: Shimizu Chikara , Kubo Mitsumasa , Kijima Hiromichi , Ishizu Akihiro , Katoh Tsutomu , Koike Takao
Published: March 2003

We treated a man with giant cell granulomatous hypophysitis with pituitary enlargement, as seen on magnetic resonance imaging. Endocrinological examination revealed panhypopituitarism and diabetes insipidus. Microscopic examination of the specimen obtained by transsphenoidal pituitary biopsy revealed a granulomatous lesion, composed of epitheliod cells, Langhans' multinucleated giant cells, lymphocytes and other chronic inflammatory cells. On whole body gallium‐67 scintigraphy, there was extensive uptake in the pituitary gland. Gallium‐67 scintigraphy may greatly aid in the diagnosis of granulomatous hypophysitis.
Reduced growth hormone (GH) responsiveness to combined GH‐releasing hormone and pyridostigmine administration in the Prader–Willi syndrome
Authors: Grugni  , Guzzaloni  , Moro  , Bettio  , De Medici  , Morabito
Published: March 2003

OBJECTIVE

It is unclear whether the blunted GH secretion in Prader–Willi Syndrome (PWS) is a true deficiency, or merely secondary to obesity. We have investigated the role of obesity in the blunted GH secretion in PWS.

DESIGN

We studied the GH response to a combined administration of GHRH (1 μg/kg i.v. at 0 min) and pyridostigmine (PD) (60 and 120 mg by mouth for children and adults, respectively, at time −60 min), as well as the baseline IGF‐I levels, in a group of patients with PWS. Two different control groups were studied with GHRH+PD using the same doses and methods as above: prepubertal and pubertal obese subjects, and prepubertal short normal children. Moreover, in 14 patients with PWS and in the group of short normals the GH response to at least two stimulation tests (insulin tolerance test, clonidine, L‐dopa, arginine) had been previously determined.

PATIENTS

Twenty‐two PWS patients (10 males and 12 females), 21 with essential obesity (11 males and 10 females), and eight short normal children (4 males and 4 females) were studied after obtaining informed consent.

MEASUREMENTS

Blood samples were taken at −60, −30 and 0 min and then 15, 30, 45, 60, 90 and 120 min after GHRH administration. Serum GH was measured in duplicate by IRMA, and IGF‐I by RIA after acid ethanol extraction. Statistical analysis was performed by t‐test for unpaired data, and analysis of variance for parametric or nonparametric data, where appropriate.

RESULTS

The GH response to GHRH+PD was significantly lower in PWS patients (AUC: mean  ± SE: 599 ± 99 μg/l/h) if compared with either short normal children (3294 ± 461 μg/l/h: P < 0.0001) or obese subjects (1445 ± 210 μg/l/h: P < 0.005). Low IGF‐I concen_trations were found in all PWS patients, so that PWS group had mean IGF‐I levels significantly lower than the other groups.

CONCLUSIONS

Our results showed that subjects with PWS had a reduced GH responsiveness to GHRH+PD associated with subnormal IGF‐I levels. These findings suggested that short stature in PWS may be at least partially correlated to the presence of GH deficiency, and that impaired GH secretion is not secondary to obesity.
Beneficial effects of long‐term GH replacement therapy on quality of life in adults with GH deficiency
Authors: Wirén Lena , Bengtsson Bengt‐Åke , Johannsson Gudmundur
Published: March 2003

OBJECTIVE

Quality of life tends to be adversely affected in adults with GH deficiency. The aim of this study was to examine changes in quality of life in a large group of GH‐deficient adults receiving long‐term GH replacement therapy.

DESIGN

The study was conducted in two stages. The first stage was a prospective, open trial of GH replacement therapy in 71 GH‐deficient adults, during which GH was administered for 20–50 months. For the second stage, a further 90 patients were recruited to create a population of 161 GH‐deficient adults, all of whom had received GH replacement therapy for more than 12 months. This population was investigated retrospectively, using a questionnaire designed specifically for this study.

PATIENTS

All patients were GH deficient and showed a peak GH response of less than 3.0 μg/l during an insulin tolerance test in which blood glucose levels dropped to 2.2 mmol/l or below.

MEASUREMENTS

The quality of life of patients in the prospective stage was assessed using the Nottingham Health Profile (NHP) parts I and II, and the Psychological General Well‐Being (PGWB) index. In the retrospective study patients completed a specially developed questionnaire, designed to determine whether changes in quality of life were sudden or gradual, and whether quality of life continued to change after GH had been administered for more than 12 months.

RESULTS

In the prospective stage, mean overall score on the NHP I improved significantly during the first 6 months of GH therapy, and remained at its new level thereafter. Scores in all areas of the NHP II improved significantly between baseline and 20–50 months, as did the total score and five of six area scores on the PGWB index. The total PGWB score and three area scores improved significantly between 6 or 12 months and 20–50 months. In the retrospective stage, 92.7% of the patients reported that they had experienced positive effects of GH therapy. In 30.3% of patients, however, such effects did not become apparent until GH had been administered for more than 6 months. Almost 60% of patients felt that their condition was still improving.

CONCLUSIONS

These results indicate that the previously reported beneficial effects of GH therapy on quality of life in GH‐deficient adults are sustained during long‐term therapy. In addition, they suggest that, once started, GH therapy should be continued for at least 6 months before judgements are made regarding its efficacy in improving quality of life.
Serum growth hormone (GH) binding protein, IGF‐I and IGFBP‐3 in patients with β‐thalassaemia major and the effect of GH treatment
Authors: Low L. C. K. , Postel‐Vinay M. C. , Kwan E. Y. W. , Cheung P. T.
Published: March 2003

OBJECTIVE

Levels of IGFI have been shown to be low in transfusion‐dependent thalassaemia and there is preliminary evidence to suggest that this may be reversed by GH treatment. In this further study we have evaluated serum growth hormone (GH) binding protein (GHBP), IGF‐I and IGFBP‐3 in patients with β‐thalassaemia major and the effects of GH treatment on these various parameters.

PATIENTS

Fifty‐six transfusion dependent patients with β‐thalassaemia major without GH deficiency between 2 and 20 years of age were studied. Thirteen non‐GH deficient patients with heights of −1.5 SD or more were treated with GH at a dose of 0.14 IU/kg/day subcutaneously for 1 year.

MEASUREMENTS

Serum GHBP, IGF‐I and IGFBP‐3 were measured in all the patients. In the 13 patients treated with GH, these serum parameters were measured before and after 3, 6 and 12 months of treatment.

RESULTS

The mean serum GHBP concentrations were normal in both prepubertal and pubertal children but the serum IGF‐I and IGFBP‐3 concentrations were low throughout childhood and adolescence. There was a significant correlation between serum IGF‐I and IGFBP‐3 concentrations (r = 0.79; P = 0.0001) but there was no correlation between the height SDS of the patients with serum GHBP, IGF‐I or IGFBP‐3 levels. GH treatment in the 13 patients resulted in significant growth acceleration associated with a significant rise in the serum IGF‐I and IGFBP‐3 and a significant fall in serum GHBP concentrations.

CONCLUSIONS

The low serum concentrations of IGF‐I and IGFBP‐3 in the presence of normal GH reserve and serum GHBP concentrations in patients with β‐thalassaemia suggest a state of partial GH insensitivity at the post‐receptor level. This partial GH insensitivity state can be overcome by supraphysiological doses of exogenous GH. The lack of correlation of IGF‐I, IGFBP‐3 and GHBP with height SDS of the patients imply that the growth failure commonly observed in patients with β‐thalassaemia major may not be specifically related to dysregulation of the GH–IGF‐I axis. GH therapy resulted in significant increase in serum IGF‐I and IGFBP‐3 but a significant fall in GHBP.
Effects of flutamide on pituitary and adrenal responsiveness to corticotrophin releasing factor (CRF)
Authors: De Leo V. , La Marca A. , Lanzetta D. , Cariello P. L. , D'Antona D. , Morgante G.
Published: March 2003

OBJECTIVE

Flutamide is a non‐steroid antiandrogen that specifically blocks the androgen receptor. We have investigated the effect of flutamide treatment on the adrenal androgen response to corticotrophin releasing factor (CRF) in eight patients with polycystic ovary syndrome (PCOS).

PATIENTS

Eight women with moderate to severe hirsutism, ranging in age from 19 to 23 years were enrolled in the study. Basal hormonal pattern showed anovulatory cycles, increased concentrations of LH, androstenedione and testosterone and increased LH/FSH ratio. A baseline ultrasound scan revealed polycystic ovaries in all patients. Each received 250 mg of Flutamide twice a day for 6 months.

MEASUREMENTS

Before treatment and at the end of the sixth month, women were evaluated for hirsutism score and a CRF test was performed to evaluate ACTH, cortisol and adrenal androgen responses.

RESULTS

Androstenedione (Δ4), DHEA‐S, 17‐hydroxyprogesterone, testosterone and free‐testosterone showed significantly reduced responses after six months of flutamide therapy whereas ACTH and cortisol response were similar to those before treatment. Clinical improvement in the degree of hirsutism was observed in all patients. The Ferriman‐Gallwey scores decreased from a mean of 22 ± 2 to 8 ± 1.5.

CONCLUSION

Flutamide induces a significant reduction in adrenal androgen response to the CRF test but not in the response of ACTH and cortisol. The finding that flutamide does not alter the pituitary‐adrenal axis shows that flutamide acts by reducing adrenal androgens. These results demonstrate that flutamide is not only effective in the treatment of hirsutism but also reduces adrenal androgen secretion.
The circadian rhythm of leptin is preserved in growth hormone deficient hypopituitary adults
Authors: Kousta Eleni , Chrisoulidou Alexandra , Lawrence Natasha J. , Al‐Shoumer Kamal A. S. , Parker Kim H. , McCarthy Mark I. , Johnston Desmond G.
Published: March 2003

OBJECTIVE

Leptin acts as a satiety factor in regulating food intake and body homeostasis, but its regulation is not well defined. Specific leptin receptors have been found in the brain and it has been hypothesized that leptin production by adipose tissue is under neuroendocrine control. A circadian rhythm has been demonstrated with highest leptin levels between midnight and early morning hours. The possibility that hypopituitarism (or pituitary surgery ± radiotherapy) abolishes this leptin rhythm was investigated by measuring serum leptin levels during a 24‐h period in patients with impaired pituitary function.

PATIENTS AND DESIGN

Circulating leptin levels were measured hourly over 24‐h in 14 hypopituitary patients (8 women and 6 men) using a sensitive and specific radioimmunoassay. Hypopituitarism was the consequence of pituitary tumors treated surgically and/or with radiotherapy. All patients were GH deficient and were receiving conventional replacement with cortisol (n = 13), thyroxine (n = 12) and desmopressin (n = 4) but not with GH.

RESULTS

A significant diurnal variation in circulating leptin concen_trations was observed in 13 of the 14 patients. The mean (±SEM) leptin levels for 8 women were 51.9 (±10.7) ng/ml and for 6 men 11.0 (±2.0) μg/l. The overall lowest leptin levels (29.3 ± 7.9 ng/ml) were observed at 0830 h after overnight fasting, rising gradually to maximum levels (43.0 ± 9.8 ng/ml) at 0200 h declining thereafter towards fasting values. The mean (±SEM) magnitude of circadian variation in absolute leptin levels from the calculated mean level for each patient was 5.6 (±1.2) ng/ml (8.4 ± 1.4 for women and 1.9 ± 0.3 for men). The mean (±SEM) of the ratio of the amplitude versus mean leptin levels over 24 h for each individual patient was 0.18 (±0.02) (0.19 ± 0.03 for women and 0.18 ± 0.02 for men).

CONCLUSIONS

A circadian rhythm for leptin is generally present in hypopituitary patients who had undergone pituitary surgery and/or radiotherapy, with the highest serum leptin levels being obtained between midnight and early morning hours. Although some patients had some residual pituitary activity, intact hypothalamic‐pituitary function is not essential for leptin's circadian rhythm.
L‐5‐hydroxytryptophan does not stimulate LH secretion directly from the pituitary in patients with gonadotrophin releasing hormone deficiency
Authors: Lado‐Abeal Joaquin , Graña Maria , Rey Carlos , Cabezas‐Cerrato Jose
Published: March 2003

OBJECTIVE

There is abundant histological and physiological evidence that serotonin plays a role in the regulation of LH secretion in rats. Studies in human subjects have been few, but their results include the finding that pulsatile administration of L‐5‐hydroxytryptophan (5‐HTP, the immediate precursor of serotonin) amplifies LH secretion in women in the medium‐late follicular phase, and that this effect is not due to 5‐HTP directly inducing LH secretion by the pituitary. We have investigated whether 5‐HTP amplifies LH secretion by enhancing the response of the pituitary to GnRH.

PATIENTS

Seven patients aged 20–40 years with hypogonadotrophic hypogonadism (HH) of hypothalamic origin (3 men with Kallmann's syndrome, 2 women without anosmia and with GH deficiency, and 2 women with anorexia nervosa).

DESIGN

To prime the pituitary, subcutaneous pulsatile GnRH was administered for 7 days at the rate of one 5–20 μg pulse every 90 min. The day before the investigation, this regimen was replaced by 1.5–3 μg intravenous pulses at the same frequency. On the day of the investigation, 3 ml blood samples were taken every 10 min from 0850 to 19.00 hours. After the first two samples, the intravenous GnRH pulse frequency was increased to one per hour and was maintained at this level throughout the rest of the study. The first 4 h of the study acted as a control phase allowing determination of the pituitary response to GnRH. At 1300 h, 75 mg of the aromatic‐L‐amino‐acid decarboxylase inhibitor carbidopa was administered orally; carbidopa does not cross the blood–brain barrier, and prevents peripheral conversion of 5‐HTP to serotonin. At 1600 h, another 75 mg dose of carbidopa was administered, and administration of 8–20 mg pulses of 5‐HTP at a rate of one pulse per hour was begun.

MEASUREMENTS

LH was determined in triplicate by an immunoradiometric assay (IRMA), and LH pulses identified by means of a program developed in our laboratory.

RESULTS

When pulsatile administration of GnRH was accompanied by administration of carbidopa and 5‐HTP, LH pulse amplitude (2.32 ± 0.71 IU/l) did not differ significantly from its value in either the GnRH + carbidopa phase (2.58 ± 1.12 IU/l) or the unaccompanied GnRH phase (2.77 ± 1.76 IU/l).

CONCLUSIONS

L‐5‐hydroxytryptophan‐induced amplification of LH secretion in humans is not due to enhancement of the pituitary response to GnRH. The effect of L‐5‐hydroxytryptophan must therefore be due to its action on the hypothalamus, where it may be hypothesized that it increases GnRH release.
Hypokalaemia during insulin‐induced hypoglycaemia in hypopituitary adults with and without growth hormone deficiency
Authors: Davies J. S. , Hinds N. P. , Millward E. M. , McDowell I. , Scanlon M. F.
Published: March 2003

OBJECTIVE

We have investigated plasma potassium changes during insulin‐induced hypoglycaemia (IIH) in adult patients with growth hormone deficiency (GHD) who have low total body potassium and may also have a vulnerable myocardium due to an increased prevalence of atherosclerosis.

DESIGN

Hypoglycaemia was induced through the administration of intravenous soluble insulin (0.1 U/Kg, IV).

SUBJECTS

23 consecutive adult patients undergoing routine biochemical evaluation for hypopituitarism.

MEASUREMENTS

GH, cortisol glucose, sodium and potassium levels were measured at 0, 30, 60, 90 and 120 min after insulin administration. Pituitary function was also assessed through measurement of TSH, thyroid hormones, LH/FSH, testosterone and 17‐oestradiol. IGF‐I concentrations were also analysed.

RESULTS

All patients achieved adequate hypoglycaemia (nadir glucose < 2 mmol/l). 15 patients had GHD (peak GH < 10 mU/l) either in isolation or as part of a spectrum of pituitary hormone deficiencies. The remaining 8 patients had normal pituitary function. Plasma potassium concentrations (mean ± SEM) fell from 3.8 ± 0.3 mmol/l (normals) and 3.8 ± 0.2 mmol/l (GHDs) to nadir concentrations of 3.0 ± 0.2 mmol/l and 3.1 ± 0.3 mmol/l, respectively (P < 0.005). Overt hypokalaemia (< 3.5 mmol/l) occurred in 13/15 GHDs and all normals. There were no significant differences between the groups.

CONCLUSIONS

Insulin‐induced hypoglycaemia causes similar degrees of significant hypokalaemia in patients with normal pituitary function and in those with GH deficiency, either alone or in combination with other pituitary hormone deficiencies. Therefore, insulin‐induced hypoglycaemia does not appear to be associated with any greater risk of hypokalaemia in hypopituitary adults with GHD compared to those with normal anterior pituitary function.
Gonadotrophin and prolactin secretory dynamics in girls with normal puberty, idiopathic precocious puberty and precocious puberty due to hypothalamic hamartoma
Authors: Uriarte M. Mercedes , Oerter Klein Karen , Barnes Kevin M. , Pescovitz Ora H. , Loriaux D. Lynn , Cutler, Jr. Gordon B.
Published: March 2003

OBJECTIVE

This study was designed to test the hypothesis that hypothalamic hamartoma causes precocious puberty through a different neuroendocrine mechanism than that of normal puberty or of idiopathic precocious puberty.

DESIGN AND PATIENTS

We compared the pattern of gonadotrophin secretion among 4 girls with precocious puberty due to hypothalamic hamartoma, 27 girls with idiopathic precocious puberty, and 14 girls with normal puberty. All subjects were breast stage 3 or 4. Blood samples were obtained every 20 min for 4 h during the day (1.000 hours to 1400 h) and night (22.00 hours to 0200 h).

MEASUREMENTS

LH, FSH, and prolactin were measured in each blood sample. Girls also underwent LHRH‐stimulation with measurement of LH and FSH before and after stimulation.

RESULTS

There were no significant differences in mean LH level, LH peak amplitude, or LH or FSH peak frequency during either the day or the night among the three diagnostic groups. However, the mean ± SD LHRH‐stimulated peak LH levels were greater in girls with hypothalamic hamartoma than in girls with normal puberty or with idiopathic precocious puberty (194 ± 142 vs 85 ± 60 or 66 ± 54 IU/l, respectively, P < 0.05). The LHRH‐stimulated peak FSH level in girls with hypothalamic hamartoma exceeded the level for the normal pubertal girls (31 ± 19 vs 17 ± 7 IU/l, P < 0.05), but not the level for the girls with idiopathic precocious puberty (25 + 12 IU/l). The peak LH to peak FSH ratio in the girls with hypothalamic hamartoma exceeded the ratio for the girls with idiopathic precocious puberty (7.3 ± 3.9 vs 2.6 ± 3.0 IU/l, P < 0.05), but not the ratio for the normal pubertal girls (5.0 + 2.9). There were no significant differences in mean prolactin level, peak amplitude or frequency, or in the ratio of mean night to mean day prolactin, among the 3 diagnostic groups.

CONCLUSIONS

We conclude that spontaneous gonadotrophin and prolactin secretion are similar among girls with hypothalamic hamartoma, idiopathic precocious puberty, or normal puberty. However, the increased LHRH‐stimulated peak LH in the girls with hypothalamic hamartoma suggests subtle differences in neuroendocrine regulation that may underlie their more rapid pubertal maturation.
The relationship between the serum leptin concentrations of thyrotoxic patients during treatment and their total fat mass is different from that of normal subjects
Authors: Zimmermann‐Belsing Tina , Dreyer Marianne , Juul Holst Jens , Feldt‐Rasmussen Ulla
Published: March 2003

OBJECTIVE

Previous studies of leptin in thyrotoxic human subjects have been short‐term and cross‐sectional. We have measured serum leptin concentrations in thyrotoxic patients in order to study the influence of endogenous thyroid hormones on the relationship between serum leptin and fat mass.

DESIGN

patients, measurements: In 10 fasting thyrotoxic patients (8 females, 2 males, mean age: 51 years) we measured serum leptin (μg/l), total thyroxine (TT4), total triiodothyronine (TT3), thyrotropin (TSH) and by Dual Energy X‐ray Absorptiometry (DEXA) total fat mass (TFM, kg) at time of diagnosis (0 months, baseline) and during 12 months treatment with thiamazole. For comparison 16 fasting thiamazole‐treated euthyroid patients (14 females, 2 males, mean age: 38 years) were studied after one year follow‐up (26 month (15–48)) and 18 normal controls (12 females, 6 males, mean age: 39 years).

RESULTS

The serum leptin concentration was 9.1 (1.3 μg/l (mean (SEM) in the thyrotoxic patients and increased significantly to 16.0 (1.3 μg/l (P < 0.0005) after 12 months treatment compared to both normal subjects and their own baseline. There was a significant correlation between serum leptin concentration and TFM in the normal control group (r = 0.79, P < 0.00009), in the thiamazole‐treated euthyroid group (r = 0.85, P < 0.00003) and in the baseline thyrotoxic group (r = 0.84, P < 0.002) but the serum leptin/TFM ratio increased significantly during 12 months of antithyroid drug treatment from 0.34 (1.2 μg/l/kg to 0.53 (1.2 μg/l/kg (P < 0.03).

CONCLUSION

The thiamazole‐treated thyrotoxic patients increased their serum leptin concentrations during 12 months antithyroid drug treatment without a significant corresponding degree of changes in TFM as expected from normal controls. It is suggested that the metabolic state in thyrotoxic patients can influence the regulation of serum leptin concentrations without any associated changes in TFM.
The effects of interleukin‐2 treatment on endothelin and the activation of the hypothalamic–pituitary–adrenal axis
Authors: Raab Claudia , Weidmann Eckhart , Schmidt Alexander , Bergmann Lothar , Badenhoop Klaus , Henning Usadel Klaus , Haak Thomas
Published: March 2003

OBJECTIVE

Recent reports suggest that complex interactions exist between the neuroendocrine and immune systems. It has been shown for example that cytokines are able to stimulate the hypothalamo‐pituitary‐adrenal axis. In addition, some studies present evidence that endothelin is able to modulate the activity of several hypothalamic‐pituitary axes, e.g. by inducing the ACTH production.

DESIGN

We investigated the effects of interleukin‐2 on endothelin levels and the hypothalamo‐pituitary‐adrenal axis. We determined the interleukin‐6, big‐endothelin, endothelin‐1, ACTH, cortisol and AVP responses to intravenously and subcutaneously administered interleukin‐2 in 8 cancer patients in a randomized placebo controlled trial.

PATIENTS

8 Patients (2 female and 6 male), age 44 ± 4.8 years, were enrolled. All patients had a World Health Organization performance status of 1 or less and a Karnofsky Index of at least 80%.

MEASURMENTS

Blood‐samples were taken before and 15, 30, 45, 60, 120, 180, 240, 300 and 360 min after interleukin‐2 injection. Cytokine serum levels and the plasma levels of big‐endothelin, endothelin, ACTH and AVP were analysed using radioimmunoassays. Cortisol was assayed by an enzyme‐linked immunosorbent assay.

RESULTS

Interleukin‐2 treatment significantly increased plasma big‐endothelin levels (P < 0.01 vs basal) and endothelin‐1 levels (P < 0.05 vs basal) within two hours and this was followed by an increase in ACTH (P < 0.01 vs basal) and cortisol (P < 0.05 vs basal) within three hours. Interleukin‐6 levels increased two hours after interleukin‐2 administration (P < 0.01 vs basal). Interleukin‐2 had no detectable effect on AVP, blood pressure or heart rate.

CONCLUSIONS

Our data demonstrate that cytokines are able to activate the human hypothalamo‐pituitary‐adrenal axis in vivo. On the basis of the observed time kinetics and in connection with previous findings from in vitro and animal models, we conclude that endothelin may be a link between cytokines and corticotrophin‐releasing hormone, most probably functioning as a cytokine‐induced neuromodulator controlling pituitary functions.