bluuebunny/crossref_metadata_2025 huggingface dataset:
Autoantibody reactive with the human general transcription factor TFIIF in sera from patients with autoimmune disorders
Authors: CAI Y , KITAJIMA S , ETOH F , KINOSHITA S , OKUBO K , HAMASAKI N
Published: November 2003

Transcription factor (TF) IIF, a heteromeric protein composed of two subunits, RAP30 and RAP74, is required for both specific initiation and elongation of mRNA synthesis by RNA polymerase II. We have identified high titre of specific autoantibodies against the RAP74 subunit of TFIIF in sera from patients with systemic lupus erythematosus (SLE) as well as those with rheumatoid arthritis (RA), periarteritis nodosa (PN), Sjögren's syndrome (SS), dermatomyositis (DM), and mixed connective tissue disease (MCTD) by Western blot or immunoprecipitation. The epitopes recognized by autoantibodies were shown to be preferentially clustered at the central charged region. Anti-RAP74 autoantibody was shown to suppress the activity of TFIIF-stimulated elongation of mRNA synthesis by RNA polymerase II. It is concluded that some patients with autoimmune disorders develop specific autoantibodies against the RAP74 subunit of TFIIF.
Anti-Ro fine specificity defined by multiple antigenic peptides identifies components of tertiary epitopes
Authors: SCOFIELD R H , ZHANG F-C , KURIEN B T , HARLEY J B
Published: November 2003

Anti-Ro (or SSA) is a clinically important autoantibody that is found in 25–40% of patients with systemic lupus erythematosus as well as an even greater proportion of patients with Sjögren's syndrome or subacute cutaneous lupus. We have studied the binding of anti-Ro sera to multiple antigenic peptides constructed from the sequence of the 60-kD Ro molecule. The results demonstrate that sera bind these peptides in solid-phase assay. Surprisingly, some of these peptides also form a precipitin line in double immunodiffusion with anti-Ro sera. Formation of lines of identity in double immunodiffusion as well as absorption studies indicate that peptides distant in the primary amino acid sequence and without shared sequence are bound by the same antibody. In addition, data from surface plasmon resonance demonstrate that peptides identified in this manner have protein–protein interactions. Thus, these techniques may identify the components of conformational epitopes.
Serum levels of IL-10, IL-15 and soluble tumour necrosis factor-alpha (TNF-α) receptors in type C chronic liver disease
Authors: KAKUMU S , OKUMURA A , ISHIKAWA T , YANO M , ENOMOTO A , NISHIMURA H , YOSHIOKA K , YOSHIKAI Y
Published: November 2003

We previously reported that the number of TNF-α-producing cells was increased in the liver of patients with type C chronic liver disease. To understand further the pathophysiology of this change, we examined serum levels of two soluble TNF receptors, TNF-αRI (p55) and -αRII (p75), and IL-10, all of which act as TNF-α buffer, and IL-15, a novel cytokine sharing many immunological activities with IL-2, using ELISA methods. We studied control individuals and patients with type C chronic liver disease, including asymptomatic hepatitis C virus (HCV) carriers with persistently normal serum ALT values, and those with chronic hepatitis (CH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Both types of sTNF-αR closely correlated with disease progression. Patients with LC and HCC had significantly elevated levels for sTNF-αRII compared with the other patient groups and controls. Serum IL-10 levels were significantly greater in all chronic liver disease groups than in controls. With respect to IL-15, the values were high in CH, LC and HCC compared with those of controls. Notably, HCC patients showed highest values for both IL-10 and IL-15, with significant differences from the other patient groups. Serial determinations revealed that interferon (IFN) treatment for CH patients resulted in the suppression of circulating IL-10 and IL-15 levels along with decrease in serum aminotransferase values. Both cytokines remained at decreased levels after cessation of therapy in patients who went into clinical and virological remission. On the other hand, treatment did not affect serum levels of sTNF-αRs. These findings indicate that serum levels of these molecules correlated with disease progress in chronic HCV infection, and that IL-10 and IL-15 may reflect the degree of inflammation in the liver. It is also suggested that both cytokines may be related to the development of HCC.
CD8+ CD28− T cells in vertically HIV-infected children
Authors: BRUGNONI D , AIRO’ P , TIMPANO S , MALACARNE F , UGAZIO A G , CATTANEO R , DUSE M
Published: November 2003

To evaluate whether vertical HIV infection interferes with the expression of CD28 on T lymphocytes, 25 HIV-infected children and 29 seroreverted children born to HIV+ mothers were studied. The percentage of CD28− cells among CD8+ T lymphocytes was higher in HIV-infected children than in controls (P < 0.001). In fact, in HIV-infected children, this percentage was elevated from the first year of life, while in healthy seroreverted children, the proportion of CD28− cells among CD8+ cells rose progressively with age (r = 0.49; P = 0.008). In HIV+ children, the CD8+ CD28−, but not CD8+ CD28+ cell proportion was significantly correlated with immunological markers of disease progression, such as CD4+ cell loss (r = −0.65; P < 0.001) and the level of in vitro spontaneous lymphocyte apoptosis (r = 0.53; P = 0.03).
Effects of rapamycin on apoptosis of rheumatoid synovial cells
Authors: MIGITA K , EGUCHI K , ICHINOSE Y , KAWABE Y , TSUKADA T , AOYAGI T , NAGATAKI S
Published: November 2003

In the present study, we investigated the effects of an immunosuppressant, rapamycin, on bcl-2 expression and the susceptibility of human rheumatoid synovial fibroblasts to Fas-mediated apoptosis. Rapamycin treatment down-regulated bcl-2 expression on rheumatoid synovial cells in a dose-dependent manner. In contrast, Fas antigen expression was not influenced by rapamycin treatment. Rapamycin treatment also enhanced the susceptibility of rheumatoid synovial cells to anti-Fas monoclonal antibody-mediated apoptosis. Our results suggest that rapamycin augments the sensitivity of rheumatoid synovial fibroblasts to apoptosis by down-regulating bcl-2 expression. This pharmacological alteration of sensitivity to apoptosis in the rheumatoid synovium may represent a new therapeutic approach for rheumatoid arthritis.
Pathogenicity of human anti-platelet factor 4 (PF4)/heparin in vivo: generation of mouse anti-PF4/heparin and induction of thrombocytopenia by heparin
Authors: BLANK M , CINES D B , AREPALLY G , ELDOR A , AFEK A , SHOENFELD Y
Published: November 2003

Heparin-induced thrombocytopenia/thrombosis (HIT) is a severe thrombotic disorder that occurs in ≈1% of patients treated with heparin. Affected patients commonly develop antibodies that recognize PF4/heparin complexes that may form on the surface of activated platelets and on the endothelium. However, it has not been established that anti-PF4/heparin antibodies are responsible for the clinical manifestations of HIT. To address this issue, we employed a recently developed model of active immunity to study the effect of IgG anti-PF4/heparin antibody in vivo. In previous studies we have shown that it is possible to induce autoimmune diseases such as systemic lupus erythematosus (SLE), anti-phospholipid syndrome (APS) or vasculitis in naive mice by active immunization with anti-DNA, anti-cardiolipin and anti-neutrophil cytoplasmic antibodies, respectively. Immunized animals develop anti-idiotypic antibodies (Ab2) and, after 2–4 months, anti-anti-idiotypic antibodies (Ab3). Ab3s generated in this manner often simulate the binding activity of Ab1 and their expression correlates with the development of specific clinical manifestations typical of the respective human disease. Based on this experience, naive BALB/c mice were immunized with IgG anti-PF4/heparin antibodies isolated from two patients with HIT. The actively immunized mice developed mouse anti-PF4/heparin antibody (Ab3). Administration of unfractionated heparin, but not low molecular weight heparin (LMWH), to the actively immunized animals induced thrombocytopenia by day 4 of drug exposure. There was no evidence of thrombosis. The results of this study support the importance of anti-PF4/heparin antibodies in the pathogenesis of HIT. Further, this model may help to elucidate the factors responsible for thrombosis as well as providing means to assess new treatment options for patients with this disorder.
Induction of proinflammatory cytokine and antioxidant enzyme gene expression following brief myocardial ischaemia
Authors: CHANDRASEKAR B , COLSTON J T , FREEMAN G L
Published: November 2003

The purpose of this study was to determine if proinflammatory cytokines are up-regulated during reperfusion following sublethal ischaemia, and whether concurrent up-regulation of antioxidant enzymes occurs. Open-chest rats were subjected to 15 min of ischaemia followed by 1 or 3 h reperfusion (R). Myocardium from the ischaemic zone showed a significantly higher (P < 0·01) generation of thiobarbituric acid-reactive substances at 1 h and 3 h R. Northern blots showed a weak signal in controls for IL-6 mRNA (0·13 ± 0·01); this was elevated to 0·68 ± 0·12 at 1 h and 0·69 ± 0·10 at 3 h R. Neither IL-1β nor tumour necrosis factor-alpha (TNF-α) were detectable in controls. IL-1β rose to 0·78 ± 0·07 at 1 h and 0·51 ± 0·08 at 3 h R, and TNF-α rose to 0·69 ± 0·10 at 1 h and 0·38 ± 0·15 at 3 h R. Western blotting showed no signals in the control, but readily detectable signals at 1 h R; these remained high (IL-6) or decreased (IL-1β and TNF-α) at 3 h R. mRNA analysis for antioxidant enzymes revealed a weak signal in controls for catalase (CAT; 0·16 ± 0·08), glutathione peroxidase (GSH-Px; 0·15 ± 0·06) and superoxide dismutase (SOD; 0·21 ± 0·05). After 1 h R, levels increased significantly for CAT (0·46 ± 0·10; P < 0·025) and GSH-Px (0·51 ± 0·13; P < 0·01), but remained similar to controls for SOD (0·26 ± 0·15). At 3 h R the mRNA levels were significantly elevated for the three enzymes (CAT 0·48 ± 0·13; GSH-Px 0·47 ± 0·10; SOD 0·54 ± 0·08). We conclude that mRNA for proinflammatory cytokines is expressed early in reperfusion, and that the proteins are present in heart tissue. Also, reperfusion is associated with rapid expression of genes for antioxidant enzymes, which may enhance reactive oxygen intermediate (ROI) scavenging.
Colchicine derivatives inhibit neopterin production in human peripheral blood mononuclear cells (PBMC)
Authors: ALTINDAG Z Z , WERNER-FELMAYER G , SAHIN G , WACHTER H , FUCHS D
Published: November 2003

Colchicine is a microtubule disrupting agent, mostly used as treatment in various kinds of inflammatory diseases such as acute familial Mediterranean fever and Behcet's disease, as well as gout. In patients with familial Mediterranean fever treatment with colchicine induces a decline of urinary neopterin concentrations which indicates a decrease of cell-mediated immune activation. In this study, we investigated a potential effect of colchicine on the T cell/macrophage system in vitro. The human myelomonocytic cell line THP-1 and PBMC were treated with colchicine or the colchicine derivative, colcemide, in the presence or absence of 250 U/ml interferon-gamma (IFN-γ) or 10 μg/ml lipopolysaccharide (LPS) for 48 h or 96 h. Colchicine and colcemide increased neopterin/protein production in unstimulated THP-1 cells, but no such effect was apparent in cells stimulated with IFN-γ. By contrast, when PBMC were treated with colchicine or colcemide a significant reduction in neopterin formation was evident in cells without and with prestimulation by IFN-γ or LPS. In parallel, reduced production of IFN-γ was observed in PBMC. These data suggest that colchicine and colcemide are able to inhibit T cell activation within the cellular immune response.
Modulation of the allergic immune response in BALB/c mice by subcutaneous injection of high doses of the dominant T cell epitope from the major birch pollen allergen Bet v 1
Authors: BAUER L , BOHLE B , JAHN-SCHMID B , WIEDERMANN U , DASER A , RENZ H , KRAFT D , EBNER C
Published: November 2003

Several in vitro and in vivo studies indicate that application of high doses of dominant T cell epitopes can induce a state of antigen-specific non-responsiveness (anergy). In the present study, we developed a murine model of an allergic immune response to Bet v 1, the major birch pollen allergen. Mice were sensitized by injection of rBet v 1 and the allergic state was proven by the presence of allergen-specific IgE and positive immediate-type skin tests to Bet v 1. In epitope mapping experiments, an immunodominant T cell epitope of Bet v 1 in BALB/c mice was identified by the use of overlapping peptides. This peptide (BV139) was subsequently employed for treatment. Two tolerization protocols were used: in one approach, the peptide was administered to naive mice before immunization (group BV139-S), in the second, already sensitized mice were treated (S-BV139). The results demonstrated that administering high doses of the dominant T cell epitope of Bet v 1 profoundly diminished T cell proliferation to the peptide in the BV139-S group, and to the peptide as well as to the whole protein in the S-BV139 group. Skin test reactivity to Bet v 1 was reduced in the BV139-S group. However, no differences in terms of specific antibody production between treated and untreated mice could be observed. This study provides evidence that administration of dominant T cell epitopes can down-regulate the allergen-specific T cell response. Proceeding on the assumption that the T lymphocyte response to allergens is crucial for the induction and maintenance of the allergic disease, a modulation of the immune response to allergens by treatment with T cell epitope peptides could represent a promising concept for immunotherapy in the future.
Anomalies of the CD8+ T cell pool in haemochromatosis: HLA-A3-linked expansions of CD8+CD28− T cells
Authors: AROSA F A , OLIVEIRA L , PORTO G , DA SILVA B M , KRUIJER W , VELTMAN J , DE SOUSA M
Published: November 2003

The present study consists of a phenotypic and functional characterization of peripheral blood T lymphocytes in a group of 21 patients with hereditary haemochromatosis (HH), an MHC class I-linked genetic disease resulting in iron overload, and a group of 30 healthy individuals, both HLA-phenotyped. The HH patients studied showed an increased percentage of CD8+ CD28− T cells with a corresponding reduction in the percentage of CD8+ CD28+ T cells in peripheral blood relative to healthy blood donors. No anomalies of CD28 expression were found in the CD4+ subset. The presence of the HLA-A3 antigen but not age accounted for these imbalances. Thus, an apparent failure of the CD8+ CD28+ T cell population ‘to expand’, coinciding with an ‘expansion’ of CD8+ CD28− T cells in peripheral blood of HLA-A3+ but not HLA-A3− HH patients was observed when compared with the respective HLA-A3-matched control group. A significantly higher percentage of HLA-DR+ but not CD45RO+ cells was also found within the peripheral CD8+ T cell subset in HH patients relative to controls. Phytohaemagglutinin (PHA) stimulation of peripheral blood mononuclear cells (PBMC) for 5 days showed: (i) that CD8+ CD28+ T cells both in controls and HH were able to expand in vitro; (ii) that CD8+ CD28− T cells decreased markedly after activation in controls but not in HH patients. Moreover, functional studies showed that CD8+ cytotoxic T lymphocytes (CTL) from HH patients exhibited a diminished cytotoxic activity (approx. two-fold) in standard 51Cr-release assays when compared with CD8+ CTL from healthy controls. The present results provide additional evidence for the existence of phenotypic and functional anomalies of the peripheral CD8+ T cell pool that may underlie the clinical heterogeneity of this iron overload disease. They are of particular relevance given the recent discovery of a novel mutated MHC class I-like gene in HH.
Calorie restriction decreases platelet-derived growth factor (PDGF)-A and thrombin receptor mRNA expression in autoimmune murine lupus nephritis
Authors: TROYER D A , CHANDRASEKAR B , BARNES J L , FERNANDES G
Published: November 2003

Calorie restriction (CR) and supplementation with fish oil (FO) are known to increase the life span and diminish histological evidence of glomerulonephritis in lupus prone (NZB×NZW)Fl (B/W) mice. Cellular proliferation is an important pathological element in the development of lupus nephritis, and we have examined the expression of thrombin receptor (TR) and the mitogenic agents PDGF-A and -B. Weanling B/W mice were fed either ad libitum or a calorie restricted (CR; 40% less calories than ad libitum) diet supplemented with either 5% (w/w) corn oil (CO) or FO. CR animals consumed 2·7–3·0 g of wet food per day versus 4·5–5·0 g for the ad libitum animals. Renal RNA was extracted from young (3·5–4·0 months of age) and old (8–10 months of age) mice. Densitometric analysis (reference gene GAPDH) of blots from Northern (PDGF-A and -B) and ribonuclease protection assays (TR) produced the following data: (i) in young mice no signal was detected for PDGF-A, -B and TR in all four groups, while the signals were readily detectable in old mice; (ii) in old mice low and similar levels of PDGF-B were detected, and neither CR nor the source of lipid altered its expression; (iii) CR significantly inhibited PDGF-A and TR expression in both CO (ad libitum versus CR; PDGF-A, 3·25-fold, P<0·025; TR, 3·7-fold, P<0·01) and FO (ad libitum versus CR; PDGF-A, 4·56-fold, P<0·01; TR, 3·6-fold, P<0·025) groups; (iv) although FO (versus CO) produced a trend towards decreased expression, results were not statistically significant. We conclude that suppression of renal disease in lupus-prone mice by CR is accompanied by decreased expression of PDGF-A and the thrombin receptor.
Enrichment of antibodies against phospholipids in circulating immune complexes (CIC) in the anti-phospholipid syndrome (APLS)
Authors: ÅRFORS L , LEFVERT A K
Published: November 2003

The aim of this study was to analyse and characterize immunoglobulins in CIC and serum from patients with anti-cardiolipin antibodies. CIC from five patients were isolated by gradient centrifugation and gel filtration. The distribution between serum and CIC of immunoglobulin reactivity against different phospholipids was determined. Serum and CIC contained antibodies against cardiolipin and other negatively charged phospholipids. The relative concentration of these antibodies was higher in the immune complexes than in corresponding sera, and the avidity of antibodies in immune complex form was higher. The presence of high concentrations of antibodies to negatively charged phospholipids in CIC from patients with anti-cardiolipin antibodies could be of pathogenic significance in APLS by conferring characteristics to the complexes of importance for binding to membrane components. This could have special implications with regard to platelet and complement activation, thrombocytopenia and thrombophilia.
Circulating levels of soluble CD30 are increased in patients with systemic sclerosis (SSc) and correlate with serological and clinical features of the disease
Authors: GIACOMELLI R , CIPRIANI P , LATTANZIO R , DI FRANCO M , LOCANTO M , PARZANESE I , PASSACANTANDO A , CIOCCI A , TONIETTI G
Published: November 2003

Activated Th2 lymphocytes express the surface molecule CD30 and release a soluble form of the same molecule which can be detected both in vivo and in vitro. In the present study, high levels of soluble CD30 were found in the peripheral blood of patients with SSc, and a significant correlation with skin score and erythrocyte sedimentation rate (ESR) was detected. Furthermore, we observed a higher spontaneous release of soluble CD30 in the supernatants of unstimulated cultures of peripheral blood mononuclear cells from our patients compared with healthy controls. Taken together, these data suggest a possible involvement of Th2 cells in the immunopathogenesis of SSc, and the dosage of CD30 soluble in the peripheral blood may be helpful in following the outcome of the disease.
Failure of plasmapheresis, corticosteroids and thionamides to ameliorate a case of protracted amiodarone‐induced thyroiditis
Authors: Samaras Katherine , Marel Geoffrey M.
Published: November 2003

We report a case of amiodarone‐induced thyrotoxicosis of protracted duration, unresponsive to conventional thionamide therapy, with therapy limited by severe adverse drug reactions. Other treatment modalities included high dose corticosteroids, plasmapheresis, lithium and perchlorate. Temporary amelioration was achieved following plasmapheresis; however, this and other measures were unsuccessful in controlling the thyrotoxicosis, which deteriorated to thyroid storm. Histopathologically, a degenerative, inflammatory thyroiditis was evident.

We discuss the limitations of conventional drug therapy and the lack of a sustained response to plasmapheresis. The failure of high dose steroids to alter the course of illness and to completely suppress the thyroidal inflammatory process is highlighted. A potential role for renal and hepatic impairment in the observed protracted course of amiodarone‐induced thyrotoxicosis is suggested.
Hypoparathyroidism and insulin‐dependent diabetes mellitus in a patient with Kearns–Sayre syndrome harbouring a mitochondrial DNA deletion
Authors: Isotani Haruhiko , Fukumoto Yoshito , Kawamura Hiroshi , Furukawa Keizo , Ohsawa Nakaaki , Goto Yu‐ichi , Nishino Ichizo , Nonaka Ikuya
Published: November 2003

We report a 17‐year‐old girl with short stature, external ophthalmoplegia, atypical retinal pigmentary degeneration, sensorineural hearing loss, and cardiac conduction defect (Kearns–Sayre syndrome). A large‐scale deletion (6741 base pairs) in mitochondrial DNA was found in her muscle specimen. She also had insulin‐dependent diabetes mellitus (IDDM). On admission, her plasma glucose level was elevated at 31.0mmol/l with mild ketoacidosis, and haemoglobinA1c elevated at 16.5%. After improvement of diabetic ketoacidosis, she was placed on insulin 24–30 units/day despite her small body weight of 25 kg. There was reduced excretion of urinary C‐peptide at 3.97 nmol/day. In addition, she had idiopathic hypoparathyroidism with a serum calcium level of 2.15 mmol/l, phosphate 1.7 mmol/l, and intact PTH below 10 ng/l. Human leucocyte associated antigen typing showed A24, A26; B54, B61; CW1, CW3; DR8, DR14; DQ1 and DQ3, suggesting that the presence of HLA‐A24 and CW3 antigen contributed to the association of IDDM and hypoparathyroidism, similar to Japanese patients with polyglandular autoimmune syndrome, complicated by hypoparathyroidism and IDDM. We suggest that a genetic linkage, as well as mitochondrial dysfunction, may be responsible for the association of the two disease states. This is an extremely rare case of Kearns–Sayre syndrome, presenting in association with IDDM and idiopathic hypoparathyroidism.
The identification of false positive responses to the pentagastrin stimulation test in RET mutation negative members of MEN 2A families
Authors: Marsh Debbie J. , McDowall Deborah , Hyland Valentine J. , Andrew Scott D. , Schnitzler Margaret , Gaskin Elizabeth L. , Nevell David F. , Diamond Terrence , Delbridge Leigh , Clifton‐Bligh Phillip , Robinson Bruce G.
Published: November 2003

OBJECTIVE The pentagastrin stimulation test is the traditional test used for the identification of asymptomatic individuals in multiple endocrine neoplasia type 2A (MEN 2A) and familial medullary thyroid carcinoma (FMTC). The identification of mutations in the RET proto‐oncogene segregating with the disease phenotype in MEN 2A and FMTC families has made it possible to re‐examine the validity of using this test for the identification of affected family members.

DESIGN Sequential and single pentagastrin stimulation test data were collected following the identification of RET mutation positive and RET mutation negative members of families with MEN 2A or FMTC.

PATIENTS RET mutations were identified in 16 Australian and New Zealand MEN 2A or FMTC families. An analysis of 39 individuals from these families was included in this study. Thirty‐two individuals (14 males, 18 females) had previously been determined as RET mutation negative. Seven individuals (6 males, 1 female) had previously been determined as RET mutation positive. Two RET mutation negative males had thyroidectomy based on prior pentagastrin test results.

MEASUREMENTS Serum calcitonin levels in response to stimulation with pentagastrin were measured at 0, 1, 2, 5 and 10 minutes post injection. Mutation analysis of the RET proto‐oncogene was performed in all individuals. In two RET mutation negative individuals from two MEN 2A families, thyroidectomy was performed and C‐cells were quantitated in order to determine the diagnosis of C‐cell hyperplasia.

RESULTS There was a statistically significant difference (P < 0.013) between RET mutation negative male and female mean peak calcitonin responses of 282 ± 236 and 96 ± 62 (mean ±SD) ng/l respectively. False positive responses to pentagastrin stimulation were identified in seven individuals who were RET mutation negative in two of the 16 families. Histologic examination of the thyroid glands in the two RET mutation negative individuals who had thyroidectomy demonstrated C‐cell hyperplasia in one but not in the other.

CONCLUSIONS There is considerable overlap between pentagastrin test results in individuals who are RET mutation positive and those who are RET mutation negative. These results indicate a need for routine performance of RET proto‐oncogene analysis on all individuals at risk of developing MEN 2A or FMTC and a coupling of pentagastrin test results and RET proto‐oncogene analysis in the decision to proceed with thyroidectomy.
Longitudinal evaluation of the development of salivary cortisol circadian rhythm in infancy
Authors: Santiago Luciano B. , Jorge Salim M. , Moreira Ayrton C.
Published: November 2003

OBJECTIVE Although an outstanding characteristic of the adrenocortical function of children and adults is its circadian rhythm, little information is available about the age of appearance of such rhythm in infancy. The main obstacle has been the ethical difficulty in obtaining serial blood samples from healthy infants. We monitored the development of cortisol daily rhythm using non‐invasive salivary cortisol determination.

DESIGN AND PATIENTS A longitudinal study of a group of 9 healthy infants was performed. Salivary samples were obtained in the morning, afternoon and night at 2, 4, 8, 12, 16, 20 and 24 post‐natal weeks from all infants.

MEASUREMENTS Cortisol was determined by RIA in 25‐μl salivary samples. Two techniques based on assay coefficients of variation were employed to characterize a normal circadian pattern of cortisol.

RESULTS Five infants (55%) established and maintained their cortisol rhythm as early as at 2, 4 and 8 weeks of age. In the remaining 4 infants the age of appearance was 12 and 20 weeks. This rhythm emerged in the group as a whole at a mean age of 8 weeks.

CONCLUSIONS Our data suggest that, in most normal infants, the development of hypothalamic–pituitary–adrenal axis circadian maturation may occur at a much earlier age than previously described.
Normal human follicle development: an evaluation of correlations with oestradiol, androstenedione and progesterone levels in individual follicles
Authors: Thierry van Dessel H. J. H. M. , Schipper Izaak , Pache Thierry D. , Van Geldorp Hans , De Jong Frank H , Fauser Bart C. J. M.
Published: November 2003

OBJECTIVE The mechanism of dominant follicle selection remains obscure.We have investigated the association between follicle diameter and follicular steroid levels in individual human ovarian follicles throughout the menstrual cycle.

DESIGN Fluid from ovarian follicles (n = 326) was obtained in vivo during surgery from 55 regularly cycling women with proven fertility. Follicles were divided into dominant (diameter > 9 mm, n = 45) and non‐dominant (diameter ≤ 9 mm, n = 281) based on ultrasound measurements.

MEASUREMENTS Fluid was assayed for oestradiol (E2), androstenedione (AD), and progesterone (P).

RESULTS Median P and E2 levels were significantly lower (P < 0.0001) and AD levels significantly higher (P = 0.03) in non‐dominant as compared to dominant follicles. In non‐dominant follicles AD (r = 0.14, P = 0.02), but not P and E2, levels were correlated to follicular diameter, and significant changes in steroid concentrations across the menstrual cycle were absent. In dominant follicles, diameter was positively correlated with P and E2 (P < 0.001) levels, and inversely correlated with AD concentrations (P = 0.01).

CONCLUSIONS Results indicate that (1) intrafollicular oestradiol concentrations rise only in follicles exceeding 9 mm in diameter and correlate with the diameter of these dominant follicles, suggesting that significant increase in aromatase enzyme activity occurs only in the dominant follicle (2) a cycle‐independent accumulation of andro‐stenedione with size occurs in non‐dominant follicles, and (3) progesterone production occurs in the largest dominant follicles only, suggesting a limited, if any, role for progesterone during follicle development.
An ovarian lipid cell tumour causing virilization and Cushing's syndrome
Authors: Elhadd T. A. , Connolly V. , Cruickshank D. , Kelly W. F.
Published: November 2003

We report a 73‐year‐old woman with rapidly developing symptoms and signs of Cushing’s syndrome and high urinary free cortisol. She was virilized and hirsute with testosterone levels which became exceptionally high. ACTH was suppressed and CT scan of the adrenals and pelvis showed no abnormality. Ultrasound scanning showed enlargement of the left ovary. Venous catheter studies suggested a left ovarian source. A 2‐cm diameter lipid cell tumour was removed from the left ovary laparoscopically. Histology was initially benign, and clinical and biochemical cure of Cushing's syndrome was rapid. However, the neoplastic nature of the ovarian tumour was demonstrated 12 months later when peritoneal metastases were detected and there was then clinical and biochemical relapse.
OVERNIGHT DEXAMETHASONE PRE‐TREATMENT IMPROVES THE PERFORMANCE OF THE LYSINE‐VASOPRESSIN TEST IN THE DIAGNOSIS OF CUSHING'S SYNDROME
Authors: Araya Patricio
Published: November 2003

OBJECTIVE  There is no endocrine test which is completely reliable for the confirmation of Cushing's syndrome and in separation of the various aetiologies. We have tested the hypothesis that overnight dexamethasone pre‐treatment should result in a better performance of the lysine‐vasopressin (LVP) test in the diagnosis of Cushing's syndrome.

STUDY DESIGN AND PATIENTS  We studied 61 subjects, including 25 pituitary‐dependent and 9 pituitary independent Cushing's (7 adrenal tumours and 2 ectopic ACTH syndromes), 18 euadrenal controls, 4 depressed subjects, and 5 cushingoid patients. The subjects received 1 mg of dexamethasone orally at 2300 h and the following morning they were given 10 IU of lysine‐vasopressin im.

MEASUREMENTS  Plasma cortisol (RIA) was measured at times −15, 0, 15, 30, 45, 60, 75, 90 and 120 minutes.

RESULTS  The dexamethasone‐modified LVP (Dx/LVP) test resulted in four patterns of cortisol response. The dexa sensitive pattern (positive suppression and negative response to LVP) was found in euadrenal subjects; the dexa insensitive pattern (negative suppression and positive response to LVP) was seen in Cushing’s disease; a non‐responsive pattern (negative suppression and negative response to LVP) was observed only in pituitary independent Cushing's; and an indeterminate pattern (positive suppression and positive response to LVP) was equivocal, being observed in 2 control subjects, 1 patient with Cushing’s disease and 1 depressed patient. In separating control subjects from Cushing's syndromes the Dx/LVP test had 88.9% sensitivity, 100% specificity and 96.2% diagnostic accuracy; when the test was used to segregate Cushing's disease from control subjects we found 96.0% sensitivity, 100% specificity and 97.7% diagnostic accuracy. The performance variables for the Dx/LVP test in separating pituitary dependent from pituitary independent Cushing's were uniformly 100%. Depressed and cushingoid subjects did not differ from control subjects in their cortisol patterns during the test. Successful removal of the pituitary microadenoma in Cushing's disease was invariably followed by a reversal of the abnormal cortisol pattern (dexa insensitive) during the test to a dexa sensitive pattern indistinguishable from that of control subjects.

CONCLUSION These results confirm our hypothesis and suggest that an improved performance of any corticotroph stimulus (oCRH, LVP, AVP or desmopressin) in the diagnosis of Cushing's syndrome should result from pre‐treatment with dexamethasone.
Acute biochemical effects of growth hormone treatment compared with conventional treatment in familial hypophosphataemic rickets
Authors: Patel L. , Clayton P. E. , Brain C. , Pelekouda E. , Addison G. M. , Price D. A. , Mughal M. Z.
Published: November 2003

OBJECTIVE Conventional treatment of familial hypophosphataemic rickets with oral phosphate and 1α‐hydroxycholecalciferol (1αHCC) does not satisfactorily correct the metabolic or physical defects of the disease and can have adverse effects, such as nephrocalcinosis. Hyperoxaluria from increased oral phosphate intake may contribute to nephrocalcinosis. Growth hormone enhances renal tubular phosphate reabsorption and 1,25‐dihydroxycholecalciferol production in normal and in GH deficient individuals, and may thus be of benefit to patients with familial hypophosphataemic rickets.

PATIENTS We have assessed the acute effects of GH on phosphate and calcium metabolism in 6 children (age 4–14 years) with familial hypophosphataemic rickets.

DESIGN Each patient served as his/her own control and received the following in a sequential non‐randomized design: conventional treatment with oral phosphate 1.0–3.4 mmol/kg/day in 3–6 divided doses and 1αHCC 18–31 ng/kg/day—no treatment—GH 0.05 mg/kg daily—GH and 1αHCC—and GH with phosphate and 1αHCC. Each treatment was given for 7 days with 7 day periods of no treatment in between.

MEASUREMENTS AND RESULTS Glomerular filtration rate, tubular maximum rate of phosphate reabsorption per litre of glomerular filtrate (TmP/GFR) and serum 1,25‐dihydroxycholecalciferol increased with GH. Mean 24‐hour plasma phosphate concentrations did not increase with GH but were higher in the treatment phases which included phosphate and 1αHCC (P = 0.002). Serum PTH was higher when GH was given in combination with phosphate and 1αHCC compared to other phases. Urine oxalate excretion did not differ between the treatment phases.

CONCLUSIONS GH seemed to partially correct the defects in renal tubular phosphate transport and 1α‐hydroxylation of 25‐hydroxycholecalciferol. We speculate that the net effect of GH treatment was an increase in body phosphate, although this was not reflected in a change in plasma phosphate. Therefore, GH in combination with 1αHCC may act as a phosphate sparing agent, permitting treatment with lower and less frequent doses of oral phosphate and reducing adverse effects such as nephrocalcinosis.
Association of HTLV‐I with autoimmune thyroiditis in patients with adult T‐cell leukaemia (ATL) and in HTLV‐I carriers
Authors: Akamine Hiromitsu , Takasu Nobuyuki , Komiya Ichiro , Ishikawa Kazuo , Shinjyo Tetsuharu , Nakachi Ken , Masuda Masato
Published: November 2003

OBJECTIVE Human T‐lymphotrophic virus type I (HTLV‐I) is a retrovirus that causes adult T‐cell leukaemia (ATL). This virus is associated with a variety of autoimmune disorders. The possible association between HTLV‐I infection and autoimmune thyroiditis has not been fully studied. We therefore evaluated anti‐thyroid peroxidase antibodies (TPOAb) and anti‐thyroglobulin antibodies (TGAb) in the sera of patients with ATL, carriers of HTLV‐I, and in healthy control subjects to investigate the possible association between such infection and autoimmune thyroiditis.

PATIENTS AND METHODS Fifty‐two ATL patients (21 males, 31 females; mean age 56.4 years) and 50 HTLV‐I carriers (18 males, 32 females; mean age 56.7 years) were studied. The control subjects were 877 healthy adults (271 males, 606 females; mean age 54.8 years) who were negative for HTLV‐I antibody. TPOAb, TGAb, thyroxine (T4) and thyrotrophin (TSH) were measured in serum using a radioimmunoassay kit.

RESULTS Positivity for thyroid autoantibodies (TPOAb and/or TGAb) was found in 21 of 52 ATL patients (40.4%), 15 of 50 HTLV‐I carriers (30.0%), and 120 of 877 control subjects (13.7%). The difference between the HTLV‐I‐infected and the control subjects was statistically significant (P < 0.005). Female control subjects had a significantly higher prevalence of positivity for thyroid autoantibodies than the males (17.3 vs 5.5%, P < 0.001). Carriers of HTLV‐I and patients with ATL of each sex showed an equally high prevalence of positivity for thyroid autoantibodies. Of the subjects who were positive for thyroid autoantibody, 7.5% of control subjects, 19.0% of ATL patients, and 40.0% of HTLV‐I carriers had hypothyroidism. A significant difference in this respect was noted between the HTLV‐I infected subjects and the control subjects (P < 0.005).

CONCLUSIONS This study demonstrates a high prevalence of positivity for thyroid autoantibodies (TPOAb and/or TGAb) in the adult T‐cell leukaemia patients and the HTLV‐I carriers. The adult T‐cell leukaemia patients and the HTLV‐I carriers each had a high prevalence of hypothyroidism. There was an association between HTLV‐I infection and autoimmune thyroiditis.
Megaoesophagus in a patient with autoimmune polyglandular syndrome Type II
Authors: Fritzen R. , Bornstein S. R. , Scherbaum W. A.
Published: November 2003

Dysphagia and vomiting are frequently present in untreated Addison's disease. These non‐specific symptoms may be due either to the metabolic disorder and myopathy or to disorders associated with Addison's disease. We describe a patient with autoimmune adrenal failure as a feature of autoimmune polyglandular syndrome (APS) type II. This patient was referred initially because of megaoesophagus. The association of megaoesophagus with Addison's disease or any of the three types of APS has not previously been described in humans. The association of megaoesophagus and adrenal failure, however, is known to occur in Allgrove's syndrome, a disease with primary manifestation in childhood characterized by adrenal failure, achalasia and alacrimia. Moreover, there are several reports on the association of megaoesophagus with adrenocortical insufficiency and other autoimmune endocrine diseases in dogs.

Vomiting and dysphagia usually resolve with hormone substitution in patients with isolated Addison's disease. In our patient these symptoms disappeared in spite of the radiological persistence of megaoesophagus, which might have been overlooked if the diagnosis of Addison's disease had been made earlier. The occurrence of megaoesophagus might be more common than previously suspected and we suggest a systematic search for similar findings in other patients with autoimmune Addison's disease, even when minor dysphagia is present.
The prevalence of non‐classic adrenal hyperplasia due to 11β‐hydroxylase deficiency among hirsute women in a Turkish population
Authors: Keleştimur Fahrettin , Şahin Yılmaz , Ayata Demet , Tutuş Ahmet
Published: November 2003

OBJECTIVE The present study was designed to determine the prevalence of 11β‐hydroxylase deficiency in adult women with hirsutism in a Turkish population.

DESIGN AND PATIENTS One hundred and twenty‐four consecutive unselected hirsute patients were studied. An ACTH stimulation test was performed in the midfollicular phase of the cycle on the patients and 20 age‐matched controls by administration of a single bolus of 0.25 mg ACTH (1–24) at 0900 h.

MEASUREMENTS Serum 11‐deoxycortisol levels were measured before, 30 and 60 minutes after ACTH injection. Basal free testosterone (fT), SHBG, cortisol and androstenedione (A) were also measured. The diagnosis of 11β‐hydroxylase deficiency has been presumed when the serum 11‐deoxycortisol response to ACTH stimulation exceeded three times the 95th percentile of controls.

RESULTS Basal hormone levels including fT and A were significantly higher in the hirsute women than in the healthy women. SHBG was significantly lower in the hirsute patients. Basal and ACTH stimulated 11‐deoxycortisol levels were found to be significantly increased in the patients compared with the controls. Eight patients (6.5%) had an 11‐deoxycortisol response higher than three times the upper normal limit.

CONCLUSIONS Using stringent diagnostic criteria, we have found that 6.5% of the hirsute women in a Turkish population could be presumed to have 11β‐hydroxylase deficiency.
The effect of a new slow‐release, long‐acting somatostatin analogue, lanreotide, in acromegaly
Authors: Al‐Maskari M. , Gebbie Jan , Kendall‐Taylor P.
Published: November 2003

OBJECTIVE Previous studies have shown that somatostatin analogues such as octreotide and lanreotide are effective in suppressing GH and IGF‐I levels in acromegaly, but the mode of administration and the frequency of injections were inconvenient for the patients. We have evaluated the effects of a new slow‐release (SR), long‐acting formulation of lanreotide, a somatostatin analogue, on clinical, biochemical and safety responses in acromegaly.

DESIGN We studied the effects of SR‐lanreotide 30mg administered intramuscularly twice or three times monthly for 6 months. Ten patients were studied, in whom acromegaly was confirmed by clinical features,mean GH>5 mU/l, and failure to suppress GH to <2 mU/l after a 75‐g oral glucose load.

MEASUREMENTS Subjective improvement in clinical symptoms of acromegaly was graded and recorded. Any adverse reactions were noted. Plasma GH levels were measured every 10min for one hour from 0830–0930h; fasting IGF‐I levels were determined at 0830h; GH, glucose and insulin responses to oral glucose loading were measured at 0,30,60,90 and 120minutes from 0930 to 1130h. Baseline measurements were carried out and repeated at 3 and 6 months. Biliary ultrasonography was performed at baseline and 6 months.

RESULTS GH levels in the10 patients decreased from 26.8±12.0 (mean SEM) to 12.7±7.0 mU/l at 3 months (P=0.04) and 9.8±5.0 mU/l at 6 months (P=0.06). Fasting IGF‐I levels decreased from 123.2±27.0 to 73.5±13.0nmol/l at 3 months (P=0.01), and increased slightly to 97.4±31.0nmol/l (P=0.05) but remained below baseline levels at 6 months. Five patients achieved good control (GH<5 mU/l) at 3months. In the remaining 5 patients the dose frequency was increased to every 10 days and one achieved good control. IGF‐I levels normalized in 3 and 5 patients at 3 and 6 months, respectively. Fasting insulin levels and peak insulin after an oral glucose load did not change significantly at 3 months but decreased from 11.7±2.0 to 7.8±3.3 mU/l (P=0.05) and 106.2±24.6 to 53.3±14.3 mU/l (P=0.04) at 6 months, respectively. There was no significant change in fasting glucose at 3 or 6 months. Most patients reported clinical improvement in acromegalic symptoms. No major adverse events were reported, but mild to moderate gastrointestinal symptoms were recorded after the initial injections. One patient developed asymptomatic gallstones at 6 months.

CONCLUSIONS This slow‐release formulation of lanreotide given either twice or thrice monthly was well tolerated, more convenient for patients, effective in controlling and alleviating the symptoms of acromegaly, as well as suppressing GH and IGF‐I levels, and had no detrimental effects on carbohydrate tolerance in acromegaly.
The 24‐hour rhythm of oxytocin and Β‐endorphin secretion in human pregnancy
Authors: Lindow S. W. , Newham A. , Hendricks M. S. , Thompson J. W. , Van Der Spuy Z. M.
Published: November 2003

OBJECTIVE Animal work suggests that maternal oxytocin secretion is influenced by the secretion of endogenous opioids in pregnancy. Spontaneous labour and pre‐labour uterine activity follow a 24‐hour rhythm the origin of which has not been explained but may be related to diurnal changes in oxytocin secretion. This study was performed to document the changes over a 24‐hour period in maternal oxytocin and Β‐endorphin secretion.

DESIGN A 4‐hourly blood profile was undertaken for a 24‐hour period.

PATIENTS Sixteen women with singleton pregnancies of more than 36 weeks gestation and 10 women with pregnancies in the mid trimester were studied.

MEASUREMENTS Blood was sampled 4‐hourly for 24 hours beginning at 1200 h. Oxytocin was measured in all patients and Β‐endorphin‐like immunoreactivity was measured in 15 patients.

RESULTS A simple index was defined for comparing night‐time levels to daytime levels for both oxytocin and Β‐endorphin. In all cases more than 36 weeks gestation the index was positive for oxytocin (night‐time levels were higher) and in all cases the index was negative for Β‐endorphin (night‐time levels were lower). In the mid trimester women all values of the index for oxytocin were positive but in the Β‐endorphin group equal numbers demonstrated a positive or a negative index.

CONCLUSIONS Reciprocal 24‐hour rhythms were demonstrated between oxytocin and Β‐endorphin; however, it is not clear whether this relationship is causal.
A prospective study of psychiatric and psychological aspects of Cushing's syndrome
Authors: Kelly William F. , Kelly Miranda J. , Faragher Brian
Published: November 2003

OBJECTIVE Cushing's syndrome is associated with psychiatric and psychological disturbances. The aim of this study was to ascertain the extent of mental illness in patients before and after treatment for Cushing's syndrome.

DESIGN AND PATIENTS Patients with Cushing's syndrome were identified for a prospective study. Control patients were selected with pituitary adenomas secreting GH or PRL. The aim was to reassess patients after Cushing's syndrome had been treated.

MEASUREMENTS Psychiatric symptoms were measured and classified using the Present State Examination (PSE), and analysed on the Catego Programme. The Hamilton Rating Scale (HRS) was used to measure depression. The Crown‐Crisp Experiential Index was used to measure common psychoneurotic symptoms (anxiety, phobia, obsession, somatic, depression and hysteria scales). The Eysenck Personality Inventory was used to assess extroversion and neuroticism. Cortisol, ACTH, and other hormones were measured by conventional methods. Parametric and non‐parametric tests were used where appropriate.

RESULTS Catego analysis of psychiatric ratings showed only 8 patients of 43 with active Cushing's syndrome (19%) were normal. Psychiatric diagnoses were obtained as follows: neurotic depression in 20 (46%), possible neurotic depression in 1 (2%), reactive depression in 6 (14%), and non‐specific neurotic symptoms in 8 (19%). Additional Catego ratings of suspected other psychoses were made for 3 patients who were also depressed. None of these 43 patients with active Cushing's syndrome had ratings of schizophrenia or mania, obsessional neurosis or pathological anxiety. In the control group 13 (87%) were normal, 1 patient with acromegaly had an anxiety state and one patient with a prolactinoma had neurotic depression.

It was possible to reassess the Present State Examination after treatment in 25 patients, when cortisol levels had been substantially reduced (to normal in 88%), the percentage rated as psychiatrically normal increased from 19 to 68 (χ2=11.7, 1 d.f., P<0.01). Hamilton Rating Scale scores for depression showed significant improvements after treatment for Cushing's syndrome (mean decrease from 9.2 to 2.4, n=36, P<0.001). Crown‐Crisp experiential index data showed significant improvements in anxiety, somatic symptoms, and depression (n=25, P<0.05). Eysenck Personality Inventory assessments showed a significant improvement in neuroticism score (n=26 P=0.016), but no significant change in extroversion (P=0.5) or lie score (P=0.6).

CONCLUSIONS Most patients with Cushing's syndrome had significant psychiatric pathology, usually depressive illness. As cortisol levels were returned to normal there were significant improvements in scores for depression and anxiety. Management of patients with Cushing's syndrome should include careful assessment of psychological and psychiatric illness.