bluuebunny/crossref_metadata_2025 huggingface dataset:
Age-related differences in cell-specific cytokine production by acutely ill Malawian patients
Authors: PETTIFORD J N , JASON J , NWANYANWU O C , ARCHIBALD L K , KAZEMBE P N , DOBBIE H , JARVIS W R
Published: March 2003

Age-related changes in human cell-specific cytokine responses to acute illness have not been well examined. We therefore evaluated age-related differences in T, B and natural killer (NK) peripheral blood lymphocyte cytokine responses of 309 acutely ill hospitalized people in Malawi, Africa, <1 month–61 years of age. We used four-colour flow cytometry and performed Wilcoxon rank sum and Kruskal–Wallis tests, Pearson (rp) and Spearman (rs) correlations, and linear and logistic regression analyses to control for human immunodeficiency virus infection (HIV) status, the percentages of lymphocytes expressing CD4, and the nature of the acute infection. The percentages of CD8− and CD8+ T cells producing induced IL-8 decreased with age (rs = −0·44 and −0·53). The percentages of T cells producing TNF-α were higher, and the percentages producing IL-10 were lower, in those ≥13 than those <13 years old (medians: 17·7 versus 10·5 and 1·4 versus 3·0, respectively). The percentages of CD8− T cells producing IFN-γ were higher and stable in those ≥1 year old compared to infants (medians: 23·5 versus 10·4); the percentages of NK producing IFN-γ were higher post-infancy and then declined to relatively low levels with increasing age. The percentages of T cells producing IL-2 were highest in those 5–<31 years old (median 5·6) and lowest in those ≥31 years old (median 1·9). The ratios of the percentages of T cells producing IL-4 to those producing IL-8 and to those producing IL-10 both increased with age. These data suggest that innate immunity, represented by NK IFN-γ production, dominates in early life. A number of shifts occur after infancy and before adolescence, including a proinflammatory shift from IL-8 to TNF-γ and a type 2 shift from IL-10 to IL-4 dominance. These findings suggest distinct age-related differences in the human response to acute illness and may be useful in directing future efforts at immunomodulatory therapies.
Neonatal dendritic cells are intrinsically biased against Th-1 immune responses
Authors: LANGRISH C L , BUDDLE J C , THRASHER A J , GOLDBLATT D
Published: March 2003

Dendritic cells (DCs) were derived from human peripheral blood monocytes or cord blood monocytes cultured in the presence of IL-4 and GM-CSF. Adult and cord DCs were observed to have comparable immature phenotypes. However, the increase in surface expression of HLA-DR and CD86 after addition of LPS was significantly attenuated in cord DCs, with CD25 and CD83 expression also markedly reduced. Cord DCs were also unable to produce IL-12p70, failed to down-regulate expression of the chemokine receptor CCR5 and induced lower levels of IFN-γ production from allogeneic naive CD4+ T cells than their adult counterparts. In contrast, the kinetics of the production of TNF-α and IL-10 in response to LPS stimulation was comparable to adult DCs. The reduced ability of cord DCs to attain a fully mature adult phenotype, and to activate naive CD4+ T cells to produce IFN-γ, suggests that they are intrinsically preprogrammed against the generation of Th-1 immune responses.
A reduction in the number of peripheral CD28+CD8+T cells in the acute phase of influenza
Authors: NABESHIMA S , MURATA M , KIKUCHI K , IKEMATSU H , KASHIWAGI S , HAYASHI J
Published: March 2003

Influenza patients show a high incidence of T lymphocytopenia in the acute phase of the illness. Since CD8+ T cells play an important role in influenza virus infection, we investigated which subset of CD8+ T cells was involved in this lymphocytopenia. CD8+ T cells from eight patients with influenza A were studied for lymphocyte count, surface marker, and intracellular IFN-γ production in the acute (days 1–3) and recovery phases (days 9–12). Total and T lymphocyte counts in the acute phase were approximately three times less than in the recovery phase; however, the CD4/8 ratio was the same in both phases. The cell count reduction in the acute phase was attributed predominantly to the CD28+ CD8+ subset, compared with the CD28− CD8+ subset. The memory/activation marker CD45RO on the CD8+ T cells was assessed. The CD28+ CD45RO− subset, a naive phenotype, was reduced significantly in number in the acute phase compared with the recovery phase. The CD28+ CD45RO+ subset, a memory phenotype, was also reduced in the acute phase, but the reduction was not statistically significant. Intracellular IFN-γ in the CD8+ subset after mitogenic stimulation was measured by flow cytometry; the percentage of CD28+ IFN-γ−/CD8+ subset in the acute phase was significantly less than in the recovery phase. These results indicated that the predominant reduction of peripheral CD8+ T cells in the acute phase of influenza was from naive-type lymphocytes, suggesting that these quantitative and qualitative changes of CD8+ T cells in influenza are important for understanding the immunological pathogenesis.
Immunological and pathological comparative analysis between experimental latent tuberculous infection and progressive pulmonary tuberculosis
Authors: ARRIAGA A K , OROZCO E H , AGUILAR L D , ROOK G A W , PANDO R HERNÁNDEZ
Published: March 2003

Mycobacterium tuberculosis produces latent infection or progressive disease. Indeed, latent infection is more common since it occurs in one-third of the world's population. We showed previously, using human material with latent tuberculosis, that mycobacterial DNA can be detected by in situ PCR in a variety of cell types in histologically-normal lung. We therefore sought to establish an experimental model in which this phenomenon could be studied in detail. We report here the establishment of such a model in C57Bl/6 × DBA/2 F1 hybrid mice by the intratracheal injection of low numbers of virulent mycobacteria (4000). Latent infection was characterized by low and stable bacillary counts without death of animals. Histological and immunological study showed granulomas and small patches of alveolitis, with high expression of tumour necrosis factor alpha (TNFα), inducible nitiric oxide synthase (iNOS), interleukin 2 (IL-2) and interferon gamma (IFNγ). In contrast, the intratracheal instillation of high numbers of bacteria (1 × 106) produced progressive disease. These animals started to die after 2 months of infection, with very high bacillary loads, massive pneumonia, falling expression of TNF-α and iNOS, and a mixed Th1/Th2 cytokine pattern. In situ PCR to detect mycobacterial DNA revealed that the most common positive cells in latently-infected mice were alveolar and interstitial macrophages located in tuberculous lesions, but, as in latently-infected human lung, positive signals were also seen in bronchial epithelium, endothelial cells and fibroblasts from histologically-normal areas. Our results suggest that latent tuberculosis is induced and maintained by a type 1 cytokine pattern plus TNFα, and that mycobacteria persist intracellularly in lung tissue with and without histological evidence of a local immune response.
Profiles of IFN-γand its regulatory cytokines (IL-12, IL-18 and IL-10) in peripheral blood mononuclear cells from patients with multidrug-resistant tuberculosis
Authors: LEE J -S , SONG C -H , KIM C -H , KONG S -J , SHON M -H , KIM H -J , PARK J -K , PAIK T -H , JO E -K
Published: March 2003

This study investigated the profiles of IFN-γ and its regulatory cytokines (IL-12, IL-18 and IL-10) in response to a purified protein derivative (PPD) antigen in peripheral blood mononuclear cells (PBMC) from 18 HIV-negative patients with multidrug-resistant tuberculosis (MDRTB), and compared them with those from 19 healthy tuberculin reactors (HTR). ELISA results showed that following stimulation with PPD, IFN-γ production was significantly reduced, whereas production of both IL-18 and IL-10 was significantly elevated in MDRTB patients compared with HTR. Three out of 18 patients with MDRTB of greater than 4 years duration showed significantly elevated IL-12 p70 production, induced by in vitro PPD stimulation of their PBMC, when compared with data from HTR. However, when taken as a group, MDRTB patients were similar to HTR in their IL-12 p70-producing capacity. IL-12 p70 protein paralleled IL-12 p40 protein expression. In addition, the production of IL-12 p40 was significantly correlated with IL-10 in all patients, but was not correlated with IFN-γ. Neutralization of IL-10 increased IL-12 p40 about twofold, but did not significantly alter IFN-γ induction in MDRTB. IFN-γ in MDRTB was highly correlated with lymphoproliferation and CD4 counts, but was not correlated with IL-12, IL-18 or IL-10 production. Our findings suggest that patients with MDRTB have dysregulated IL-12, IL-18 and IL-10 production during Mycobacterium tuberculosis infection, and the cytokine profiles are similar to those in patients with drug-sensitive advanced TB previously reported in the literature. In addition, IL-10 may not have a dominant role in defective IFN-γ production in patients with MDRTB.
The transcriptional response of human macrophages to murabutide reflects a spectrum of biological effects for the synthetic immunomodulator
Authors: GOASDUFF T , DARCISSAC E C A , VIDAL V , CAPRON A , BAHR G M
Published: March 2003

The synthetic immunomodulator murabutide (MB) presents multiple biological activities with minimal toxicity in animals and in man. Although MB is known to target cells of the reticuloendothelial system and to regulate cytokine synthesis, the molecular mechanisms underlying several of its biological effects are still largely unknown. In an effort to define cellular factors implicated in the immunomodulatory and HIV-suppressive activities of MB, we have undertaken profiling the regulated expression of genes in human monocyte-derived macrophages (MDM) following a 6-h stimulation with this synthetic glycopeptide. Oligonucleotide microarray analysis was performed on RNA samples of differentiated MDM from four separate donors, using probe sets corresponding to 1081 genes. We have identified, in a reproducible fashion, the enhanced expression of 40 genes and the inhibition of 16 others in MB-treated MDM. These regulated genes belonged to different families of immune mediators or their receptors, transcription factors and kinases, matrix proteins and their inhibitors, ion channels and transporters, and proteins involved in cell metabolic pathways. Additional verification of the regulated expression of selected genes was carried out using Northern blots or the quantification of released proteins in MDM cultures. The profile of MB-regulated genes in MDM provides a molecular basis for some of its previously reported biological activities, and reveals new set of genes targeted by the immunomodulator suggesting potential application in novel therapeutic indications.
T cell receptor usage in patients with non-progressing HIV infection
Authors: BODMAN-SMITH M D , WILLIAMS I , JOHNSTONE R , BOYLSTON A , LYDYARD P M , ZUMLA A
Published: March 2003

It is still unclear why some patients with HIV progress more slowly than others to developing full blown AIDS. In this study using flow cytometry we have investigated the TCRBV repertoire of peripheral blood T lymphocytes in 17 long-term non-progressing HIV patients (LTNP) to determine if there is a biased usage of T cell receptor V gene products. Patients were identified from hospital records and entered into the study. Three colour flow cytometry was used to determine the expression of the TCRBV3S5, BV5S1, BV5S2, BV5S3, BV6S1, BV7S1, BV9, BV11, BV12, BV13, BV14, BV16, BV17, BV18, BV20, BV21S3, BV22, and BV23 by CD8 and CD4 positive cells isolated from the peripheral blood of patients and controls. Increases in the absolute numbers of CD8+ T cells expressing TCRBV2 and 8 were observed in the HIV-LTNP population (P < 0·05 in both cases). No differences were seen in numbers of CD8+ T cells expressing other TCRBV or in any TCRBV within the CD4+ T cell popu-lation. At follow up (1–2 years later), those patients in which CD4 levels were below 500 × 106/l were those initially found to have lower levels of TCRBV8 +ve CD8 cells. A significant increase in the absolute numbers of T cells coexpressing the gamma delta (γδ) T cell receptor and CD8 were also seen in the HIV-LTNP patients compared with controls (P = 0·002). The increase in CD8+ T cells in the HIV-LTNP patients may be interpreted as either an antigen specific, or group of antigen specific responses to viral antigen, or less likely a viral superantigen. A low level of TCRBV8, CD8+ T cells might be predictive of a more rapid disease progression and might indicate a protective role for this population in HIV infected patients. The increase in γδT cells bearing the CD8 coreceptor suggests a role for this cell type in the response to HIV infection.
Abnormal expression of intracellular cytokines and chemokine receptors in peripheral blood T lymphocytes from patients with systemic sclerosis
Authors: FUJII H , HASEGAWA M , TAKEHARA K , MUKAIDA N , SATO S
Published: March 2003

In patients with systemic sclerosis (SSc), there are conflicting findings regarding which is predominant between type 1 and type 2 immune responses. To determine the balance between type 1 and type 2 T lymphocytes in peripheral blood from SSc patients, we investigated the expression of intracellular cytokines, such as interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4, and IL-13, and chemokine receptors such as CXCR3 and CCR4 by flow cytometry. The frequency of IFN-γ-producing cells among CD8+ cells was significantly increased in patients with diffuse cutaneous SSc (n = 11, P <  0·0001) and limited cutaneous SSc (lSSc; n= 16, P < 0·0001) compared with normal controls (n = 17) while there was no significant difference in the frequency of IL-4- or IL-13-producing cells. In contrast, the frequency of IFN-γ- or IL-4-producing cells among CD4+ cells was similar between the three groups. Similar results were obtained when absolute numbers were assessed. The frequency of IFN-γ-producing cells among CD8+ cells inversely correlated with percentage DLco in SSc patients (r = − 0·650, P < 0·005). CXCR3+ CD8+ cells selectively produced IFN-γ, and the frequency of CXCR3+ CD45RO+ cells among CD8+ cells was higher in lSSc patients (n = 14, P < 0·01) than in normal controls (n = 22). In contrast, there was no significant difference in the frequencies of CXCR3- or CCR4-expressing CD45RO+ cells among CD4+ cells. These results demonstrate the predominance of type 1 cytokine-producing cells (Tc1 cells) in peripheral blood CD8+ T cells from SSc patients, but no definite Th1/Th2 imbalance in CD4+ T cells. Tc1 cells may be associated with pulmonary vascular damage in SSc.
Evidence for lack of cross-genotype protection of CD4+ T cell responses during chronic hepatitis C virus infection
Authors: HARCOURT G C , LUCAS M , GODKIN A J , KANTZANOU M , PHILLIPS R E , KLENERMAN P
Published: March 2003

CD4+ T lymphocyte responses are thought to play a major role in control of the hepatitis C virus (HCV). Few, however, have been mapped down to the level of peptide and HLA restriction. Furthermore, the ability of such T cells to respond to viruses which differ in genotype has not been addressed in detail. In most cases of persistent infection with HCV, CD4 proliferative responses are weak or absent. From a large cohort of persistently infected patients, we identified an individual with unusually robust and persistent responses in the face of chronic infection. We firstly mapped two peptide epitopes to regions of the nonstructural protein NS4 (aa1686–1705 and aa 1746–1765). However, in contrast to the genotype 1a derived antigens used for mapping, the infecting virus was identified as genotype 3a. Strikingly, the patient's CD4 response to these epitopes were specific only for the genotype 1a sequence, and did not recognize genotype 3a synthetic peptides. Serologic assays indicated that prior exposure to HCV of genotype 1 had occurred. This patient therefore maintains strong CD4 proliferative responses which are genotype specific and not cross-reactive. The apparent ‘misdirection’ of these nonprotective responses has important implications for the role of natural and vaccine induced CD4 responses in the face of variable viruses.
Autoantibodies against complement receptor 1 (CD35) in SLE, liver cirrhosis and HIV-infected patients
Authors: SADALLAH S , HESS C , TRENDELENBURG M , VEDELER C , LOPEZ-TRASCASA M , SCHIFFERLI J A
Published: March 2003

The acquired loss of CR1 (CD35) on erythrocytes in specific autoimmune diseases and chronic infections may be due to autoAb against CR1. An ELISA using rCR1 was established to measure antiCR1 IgG autoAb. Plasma containing alloAb to polymorphism on CR1 (Knops blood group Ab) reacted strongly against rCR1 and were used as positive controls. AntiCR1 Ab was found in 3/90 (3·5%) plasma samples from healthy blood donors. The binding of these Ab was not inhibited by high salt concentrations. AntiCR1 Ab were present in the IgG fractions of plasma, and they bound to rCR1 on Western Blot. Affinity chromatography on rCR1-sepharose depleted the plasma of antiCR1, and the acid-eluted fractions contained the antiCR1 Ab. An increased frequency of antiCR1 autoAb was found in patients with SLE (36/78; 46%), liver cirrhosis (15/41; 36%), HIV infection (23/76; 30%) (all P < 0·0001), and in patients with anticardiolipin Ab (4/21; 19%, P < 0·01) multiple sclerosis (7/50; 14%, P < 0·02), and myeloma (autoAb (8/56; 14%, P < 0·02), but not in those with acute poststreptococcal glomerulonephritis (1:32; 3%). Because C1q binds to CR1, antiC1q Ab were analysed in the same patients. There was no correlation between levels of antiC1q and antiCR1 autoAb. In HIV patients, levels of antiCR1 did not correlate with low CR1 levels expressed on erythrocytes or soluble CR1 in plasma.

The binding of antiCR1 autoAb to rCR1 fixed on ELISA plates was not inhibited by soluble rCR1 or by human erythrocyte CR1, in contrast to alloAb and one SLE serum, which induced partial blockade. Thus, antiCR1 autoAb recognize mostly CR1 epitope(s) not present on the native molecule, suggesting that they are not directly involved in the loss of CR1. Rather antiCR1 autoAb might indicate a specific immune response to denatured CR1.
Monocyte-derived cytokines in multiple sclerosis
Authors: FILION L G , GRAZIANI-BOWERING G , MATUSEVICIUS D , FREEDMAN M S
Published: March 2003

MS is an inflammatory, presumably autoimmune, disease mediated by the activation of T cells, B cells and monocytes (MO). Inflammation is thought to occur early during the relapsing-remitting phase of MS (RRMS), whereas in the later phases of MS such as secondary progressive MS (SPMS), inflammation tends to diminish. Our objective was to compare the types and amounts of proinflammatory and regulatory cytokines produced by MO from relapsing–remitting patients with or without treatment with IFN-β (RRMS+ therapy, RRMS− therapy), respectively, from secondary progressive patients (SPMS) and from healthy controls (HC). MO were isolated by a density-gradient technique and three different techniques (RNase protection assay, ELISA and intracellular cytokine staining) were used to assess cytokine levels. An increase in IL6, IL12 and TNF-α was observed by all three methods for RRMS– therapy and for SPMS patients compared to HC and RRMS+ therapy patients. We conclude that proinflammatory and regulatory monokines can be derived from MO of MS patients and that these levels are modulated by IFN-β therapy. Although it is believed that inflammation tends to diminish in SPMS patients, our data show that inflammatory cytokines continue to be released at high levels, suggesting that IFN-β or IL10 treatment may be beneficial for this group.
Antigens shared by malignant plasma cells and normal B cells may be involved in graftversusmyeloma
Authors: HOLLOWAY P A , KALDENHOVEN N , KOK-SCHOEMAKER H M , VAN KESSEL B , VAN BLOKLAND W T M , BLOEM A C , LOKHORST H M
Published: March 2003

Cytotoxic T cells play an important role in graft-versus-host-disease (GvHD) and graft-versus-leukaemia/myeloma, which may occur in patients treated with an allogeneic stem cell transplantation (ASCT). Here, we describe the selection of a myeloma reactive CD4+ cytotoxic T cell-line (CTL) and two CD4+ clones from this CTL. The CTL was generated from the blood from a patient with multiple myeloma (MM) with graft versus myeloma/GvHD, following an ASCT. The CTL was stimulated using irradiated peripheral blood mononuclear cells and EBV transformed B cells from the myeloma patient (EBVp), both of which were obtained prior to ASCT. Both the CTL and the two T cell clones specifically lysed EBVp and secreted IFN-γ after coculture with EBVp and autologous myeloma tumour cells in a class II restricted fashion. These results show that myeloma tumour cells and autologous B cells present a common polymorphic peptide that functions as a target for graft derived cytotoxic T cells. Identification of these proteins will give insight into the relationship between graft versus myeloma (GvM) and GvHD and may provide immunotherapeutical targets in the treatment of MM.
Flow cytometric determination of glucocorticoid receptor (GCR) expression in lymphocyte subpopulations: lower quantity of GCR in patients with post-traumatic stress disorder (PTSD)
Authors: GOTOVAC K , SABIONCELLO A , RABATI´C S , BERKI T , DEKARIS D
Published: March 2003

Assessment of the intracellular glucocorticoid receptor (GCR) level may be useful in monitoring functional disturbances of the hypothalamic-pituitary-adrenocortical axis or effects of prolonged steroid therapy. Cytosolic ligand binding assays have recently been supplemented by flow cytometric determination of receptor expression in individual cells. A method based on multiparametric analysis of whole blood by simultaneous labelling of intracellular GCRs and surface markers of lymphocyte subsets is described. We examined 25 healthy male volunteers and 35 age- and sex-matched post-traumatic stress disorder (PTSD) patients within 8 years from traumatic event. PTSD patients had a lower relative quantity of GCR in all lymphocyte populations tested as compared with healthy volunteers. NK cells of both groups showed higher expression of GCR than other lymphocyte subsets. In PTSD patients, the expression of GCR in B lymphocytes was also higher than in T cell. Although serum cortisol level was lower in PTSD patients, there was no correlation between cortisol level and GCR expression. Multiparameter flow cytometric determination of GCR expression in lymphocyte subpopulations may provide a useful tool for monitoring immunoregulatory action of glucocorticoids.
Enhanced chemokine response in experimental acuteEscherichia colipyelonephritis in IL-1β-deficient mice
Authors: HERTTING O , KHALIL A , JAREMKO G , CHROMEK M , LI Y-H , BAKHIET M , BARTFAI T , TULLUS K , BRAUNER A
Published: March 2003

The aim of the present study was to investigate the effects of IL-1β and Escherichia coli on the expression and secretion of MIP-2, the mouse equivalent to human IL-8, MCP-1 and RANTES in the kidneys of mice with acute pyelonephritis. Female Bki NMRI, as well as IL-1β deficient mice and their wild-type littermates, were transurethrally infected with either E. coli CFT 073 or injected with NaCl 0·9% (w/v) and thereafter obstructed for 6 h. The Bki NMRI mice were killed at 0, 24, 48 h and 6 days and the IL-1β-deficient mice at 48 h. Chemokine mRNA and protein levels peaked at 24 h for the tested chemokines with the mRNA expression localized in the tubular epithelial cells and for MIP-2 also in neutrophils. Obstruction per se, also induced a chemokine expression similar to E. coli infection although at a lower level. Interestingly, MIP-2 levels were higher in the IL-1β deficient mice as compared with the wild-type littermates. Likewise, the inflammatory changes were more frequent and, when present, more widespread in the IL-1β-deficient mice than in the wild-type mice. Stimulation of a human renal tubular epithelial cell line (HREC), A498 and of primary human mesangial cells (HMC) with the same bacterial antigen depicted gene expression of the same chemokines. A rapid release of IL-8 and MCP-1 was observed from both cell types. RANTES response was delayed both in the HREC and the HMC. We conclude that acute E. coli pyelonephritis induces a MIP-2/IL-8, MCP-1 and RANTES expression and secretion localized primarily to the epithelial cells and that this production is confirmed after in vitro stimulation with the same bacterial antigen of human epithelial and mesangial cells. Blockade of induction of chemokine response may thus be an attractive target for possible therapeutic intervention.
Prognostic factors for survival and for biochemical cure in medullary thyroid carcinoma: results in 899 patients
Authors: Modigliani Elisabeth , Cohen Régis , Campos José‐Marie , Conte‐Devolx Bernard , Maes Béatrice , Boneu Andrée , Schlumberger Martin , Bigorgne Jean‐Claude , Dumontier Philippe , Leclerc Laurence , Corcuff Bernard , Guilhem Isabelle
Published: March 2003

BACKGROUND

Prognostic factors of sporadic or inherited medullary thyroid carcinoma (MTC) are still controversial and have been assessed in old and small series. A better knowledge of these factors would improve patient management.

OBJECTIVE

To evaluate factors involved in the prognosis of MTC in a large series of cases, using uni‐ and multivariate analysis.

DESIGN AND PATIENTS

Clinical, biological, surgical and epidemiological data on 899 MTC patients, diagnosed between 1952 and 1996, were collected by the French Calcitonin Tumors Study Group (GETC) with a standardized questionnaire, and processed in a national database.

MEASUREMENTS

Survival and biochemical cure (i.e. normal basal post‐operative serum calcitonin levels) were analysed with Kaplan and Meier and log‐rank test statistical procedures. Data are presented as adjusted rather than observed survival, to consider only patients who died of MTC. Cox's forward‐stepping proportional hazard model was used to analyse factors with a significant influence on survival by univariate analysis.

RESULTS

Apart from the large proportion of familial forms (43%), the general characteristics of our population were similar to those in other studies: mean age at surgery = 43.4 years; sex ratio = 1 male/1.35 female; stage I = 20.8%; stage II = 21.2%; stage III = 46.5% and stage IV = 11.5%. 863 (96%) patients underwent surgery; 43% of operated patients were biochemically cured. Adjusted survival was 85.7 ± 1.5% at 5 years and 78.4 ± 2.1% at 10 years. Multivariate analysis showed that age and stage were independent predictive factors of survival. Gender, type of surgery, type of familial form were predictive only in univariate analysis. Biochemical cure predicts a survival rate of 97.7% at 10 years. Authentic recurrence, that is subsequent elevation of calcitonin (CT) after post‐operative normalization, was found in 4.9%. In non‐cured patients (57%), survival was still good: 80.2% (±2.2%) and 70.3% (±2.9%) at 5 and 10 years, respectively. Similarly, prediction of biochemical cure was solely dependent on stage.

CONCLUSION

Survival of these medullary thyroid carcinoma patients appears better than expected even in non‐cured patients. Considering the strong impact of stage, the necessity for pre‐operative diagnosis of MTC is obvious.
Long‐term follow‐up of bone mineral density and bone metabolism in transsexuals treated with cross‐sex hormones
Authors: Van Kesteren Paul , Lips Paul , Gooren Louis J. G. , Asscheman Henk , Megens Jos
Published: March 2003

OBJECTIVE

It is unknown whether long term cross‐sex hormone treatment affects the human skeleton. We monitored bone mineral density and biochemical markers of bone turnover for 28–63 months in 20 male‐to‐female transsexuals (M → F) treated with anti‐androgens and oestrogens, and 19 female‐to‐male transsexuals (F → M) treated with androgens. They underwent gonadectomy 13–35 months after the start of cross‐sex hormone administration.

DESIGN

Bone mineral density (BMD) and the markers of bone turnover osteocalcin, alkaline phosphatase, fasting urinary calcium/creatinine and hydroxyproline/creatinine, were measured at baseline, after 1 year and after 28–63 months of cross‐sex hormone administration.

RESULTS

In oestrogen‐treated M → F, variables of bone turnover decreased significantly with consecutive measurements. BMD had increased significantly after 1 year, but decreased again to baseline levels after 28–63 months of cross‐sex hormones.

In F → M, alkaline phosphatase levels increased during the first year. BMD did not change during the first year but had decreased significantly after 28–63 months following ovariectomy. In both M → F and F → M, the change of BMD correlated inversely with serum LH and FSH levels. Of all biochemical variables LH levels appeared to be the best predictor of loss of BMD; in the long‐term LH levels were more elevated in testosterone‐treated F → M than in oestrogen‐treated M → F transsexuals.

CONCLUSION

In M → F, oestrogen treatment prevented bone loss after testosterone deprivation. In F → M the testosterone dosage used, associated with a decline in serum oestradiol levels, was unable to maintain bone mass fully in all subjects in the longer term. The inverse relationship between BMD and serum LH levels suggests that the dose of hormone replacement has been too low in subjects with a decline in their BMD. Its cause might be underdosing or non‐compliance in some patients. We propose that serum LH levels may be used as a measure of the adequacy of replacement with sex steroids.
Tuberculous Addison's disease: lack of normalization of adrenocortical function after anti‐tuberculous chemotherapy
Authors: Bhatia Eesh , Jain Sunil K. , Gupta Rakesh K. , Pandey Rakesh
Published: March 2003

OBJECTIVE

Tuberculosis of the adrenal glands is a common cause of Addison's disease in developing countries. We conducted a study to determine if treatment of such patients with modern anti‐tuberculous chemotherapy would lead to an improvement in plasma cortisol and aldosterone levels.

DESIGN

Prospective study.

PATIENTS

5 patients with Addison's disease secondary to tuberculosis.

MEASUREMENTS

Basal and ACTH stimulated plasma cortisol and aldosterone levels were measured prior to instituting anti‐tuberculous chemotherapy, as well as one month after its conclusion. Four patients were again studied over the next 2–5 years.

RESULTS

Peak plasma cortisol levels prior to treatment were markedly reduced (range, < 14–110 nmol/l). There was no improvement one month (< 14–143 nmol/l) or 2–5 years (< 14–69 nmol/l) after completing anti‐tuberculous chemotherapy. Peak plasma aldosterone at diagnosis was < 56–210 pmol/l; it was undetectable in 4 patients. No improvement was observed one month (< 56–210 pmol/l), or 2–5 years (< 56–389 pmol/l) after stopping anti‐tuberculous chemotherapy. Plasma aldosterone levels at both these time points were far lower than those in control subjects (median 763 pmol/l, 560–1512 pmol/l; p < 0.01). One patient had an increase in peak aldosterone from < 56 pmol/l to 389 pmol/l, though peak cortisol actually declined in this subject (from 110 nmol/l to 69 nmol/l).

CONCLUSIONS

Treatment of tuberculous Addison's disease with anti‐tuberculous chemotherapy does not lead to normalization of ACTH stimulated plasma cortisol or aldosterone levels during the 2–5 year period of study. However, prolonged follow up with regular adrenal function tests is warranted in all such patients.
Radioiodine treatment in unsuspected pregnancy
Authors: Evans  , Webster  , Evans  , Bevan  , Scanlon
Published: March 2003

OBJECTIVE

Radioiodine is being used increasingly as first line therapy for hyperthyroidism. Our aim is to highlight some of the difficulties which can occur following the use of 131I to treat hyperthyroidism in fertile women.

PATIENTS

We present 3 cases of young women to whom radioiodine was given, only to find some weeks later that they had been pregnant at the time of treatment.

CONCLUSIONS

These cases serve as a reminder of the importance of obtaining an accurate and full menstrual and contraceptive history. Guidelines advocate the application of the ten day rule with the further recommendation that pregnancy testing may be undertaken as an alternative.
Serum follistatin concentrations are increased in patients with septicaemia
Authors: Michel Uwe , Shintani Yasumi , Nau Roland
Published: March 2003

OBJECTIVE

Follistatin (FS) is the specific binding protein of activin, a growth and differentiation factor of many cell types. Both factors have almost ubiquitous tissue distributions. In vitro, FS is secreted by vascular endothelial cells and this can be stimulated by bacterial compounds. For this reason, serum FS levels were examined in patients with septicaemia.

PATIENTS

Five male and four female patients of different age with various forms of septicaemia and different clinical outcome.

MEASUREMENTS

Serum concentrations of FS, C‐reactive protein (CRP) and blood leucocyte counts were determined repeatedly in all nine patients; samples from age‐ and sex‐matched healthy volunteers served as controls.

RESULTS

The median of the maximum FS concentrations of septicaemic patients was threefold higher than in healthy controls (P = 0.008). The highest increase observed was approximately 40 times normal. Serum FS levels in patients with septicaemia showed large variations between individuals. Serum FS levels parallelled those of CRP but were not correlated to the leucocyte counts.

CONCLUSION

serum follistatin concentrations in humans are elevated during septicaemia and appear to parallel serum C‐reactive protein levels.
Low level of glucocorticoid receptor messenger ribonucleic acid in pituitary adenomas manifesting Cushing's disease with resistance to a high dose‐dexamethasone suppression test
Authors: Mu Yi‐Ming , Takayanagi Ryoichi , Imasaki Kyosuke , Ohe Kenji , Ikuyama Shoichiro , Yanase Toshihiko , Nawata Hajime
Published: March 2003

OBJECTIVES

The overnight 8‐mg dexamethasone suppression test is often used to differentiate Cushing's disease, due to an oversecretion of ACTH from the pituitary gland, from other kinds of Cushing's syndrome. However, a few patients with ACTH‐producing pituitary adenoma show no suppression of plasma cortisol after the administration of 8 mg of dexamethasone. To clarify the relationship between the level of glucocorticoid receptor (GR) in the pituitary adenoma and the sensitivity to dexamethasone in Cushing's disease, we thus examined the levels of GRα and GRβ mRNAs in the pituitary adenomas in six patients who were proven at surgery to have pituitary ACTH‐producing adenomas.

MATERIALS

Total RNA was extracted from six pituitary adenomas and pituitary tissue adjacent to one of the adenomas, and the mRNA levels of GRα, GRβ, pro‐opiomelanocortin (POMC) and β‐actin in these samples were sampled by quantitative RT–PCR.

RESULTS

The GRα mRNA levels in the adenomas from the two patients who showed no response to the 8‐mg dexamethasone suppression test were significantly lower than those in the adenomas of four patients who showed suppression. The GRβ mRNA level was much lower than that of GRα mRNA but not significantly different among the six adenomas.

CONCLUSIONS

These results suggest strongly that decreased expression of GRα in pituitary adenomas may be the major reason for the marked insensitivity to the 8‐mg dexamethasone suppression test observed in two patients with Cushing's disease.
The efficacy of DDAVP is related to the circadian rhythm of urine output in patients with persisting nocturnal enuresis
Authors: Hunsballe J. M. , Hansen T. K. , Rittig S. , Pedersen E. B. , Djurhuus J. C.
Published: March 2003

OBJECTIVE

Desmopressin may be a useful treatment in some, but not all, patients with nocturnal enuresis. We have evaluated a relation between nocturnal urine output in patients with primary monosymptomatic nocturnal enuresis and the treatment response to synthetic vasopressin.

DESIGN

Adolescent or adult enuretics and normal subjects were enrolled in the study and admitted to hospital for a 24 hour investigation of the diurnal variation in urine output, plasma vasopressin (AVP) and plasma atrial natriuretic peptide (ANP). The enuretics were characterized prior to investigation as either 1‐desamino‐8‐D‐arginine vasopressin (DDAVP) responders or non‐responders. During admission the fluid intake was restricted to 25 ml/kg per day.

PATIENTS

Twenty‐four patients (15–37 years) with primary monosymptomatic nocturnal enuresis and 9 normal subjects (24–31 years).

MEASUREMENTS

Circulating levels of AVP, ANP, plasma electrolytes and plasma osmolality were measured (1400, 2000, 2300, 0200, 0500 and 0800 hours) together with urine volume, urine osmolality and urine electrolytes during daytime and nighttime. Tubular reabsorptive capacity for water, osmoles and creatinine were assessed as well as urinary and fractional excretion rates of sodium and potassium.

RESULTS

Controls and DDAVP non‐responders had a significant decrease in urine output at night concomitant with a significant plasma AVP amplitude in peak/nadir values although both groups lacked a significant nocturnal increase in AVP. In contrast, in DDAVP responders there was no circadian variation in urine output and thus a nocturnal polyuria together with no oscillation in plasma AVP. The DDAVP responding group had a nocturnal urine production significantly larger than the two other groups. However, the mean 24 hour AVP levels were similar in all groups. The excessive urine production at night in DDAVP responders was accompanied by nocturnal natriuresis due to an increased fractional excretion of sodium. In contrast, nocturnal antidiuresis in controls and DDAVP non‐responding enuretics coincided with diminished sodium excretion. Average ANP levels were elevated in both enuretic groups compared to normals, whereas a circadian variation was detected only in the latter.

CONCLUSION

It is concluded that DDAVP responsiveness is linked to the nocturnal urine production and that no pathophysiological role can be ascribed to AVP or ANP in DDAVP refractory adolescent and adult enuretics. Moreover, it is suggested that an abnormal tubular handling of sodium may contribute to the nocturnal polyuria seen in DDAVP responders.
Effects of growth hormone treatment on serum levels of insulin‐like growth factors (IGFs) and IGF binding proteins 1–4 in postmenopausal women
Authors: Kassem M. , Brixen K. , Mosekilde L. , Blum W. F. , Flyvbjerg A.
Published: March 2003

BACKGROUND AND OBJECTIVE

Insulin‐like growth factor binding proteins (IGFBPs) modulate the actions and bioavailability of insulin‐like growth factors (IGFs), however, their regulation in vivo is incompletely understood. In this study we investigated the effects of different doses of growth hormone (GH) on circulating levels of IGFs and IGFBPs.

DESIGN

The study was double‐blind and placebo‐controlled. Patients were treated with either GH in doses of 0.05, 0.10, or 0.20 IU/kg/day of placebo for one week.

PATIENTS

Forty post‐menopausal women aged 52–73 years with low bone mass.

MEASUREMENTS

Serum IGF‐I and IGF‐II were measured by RIA while IGFBP‐1–3 were measured by Western ligand blot (WLB) and compared with determinations by specific immunoassays. IGFBP‐4 was measured by WLB alone.

RESULTS

Both IGF‐I (P < 0.001) and IGF‐II (P < 0.01) increased significantly during GH treatment. Additionally, IGFBP‐1 (P < 0.001) and IGFBP‐2 (P < 0.001) decreased significantly while IGFBP‐3 (P < 0.001) and IGFBP‐4 (P < 0.05) increased all in a dose‐dependent manner. Stepwise (backwards) multiple regression analyses showed that the changes in IGF‐I and IGF‐II, and age correlated with the change in serum IGFBP‐1. Both GH‐dosage, the increase in IGF‐II, and body mass index correlated with the decrease in IGFBP‐2. Furthermore, the increase in serum IGF‐I, IGF‐II, and triiodothyronine correlated with the increase in IGFBP‐3. Moreover, GH‐dosage correlated with the increase in serum IGFBP‐4.

CONCLUSION

GH significantly increased serum IGF‐I, IGF‐II, IGFBP‐3, and IGFBP‐4 and decreased serum IGFBP‐1 and IGFBP‐2 in post‐menopausal women.
Serum oestradiol and oestrogen‐receptor gene polymorphism are associated with bone mineral density independently of serum testosterone in normal males
Authors: Ongphiphadhanakul B. , Rajatanavin R. , Chanprasertyothin S. , Piaseu N. , Chailurkit L.
Published: March 2003

OBJECTIVES

The physiological effects of oestrogens on bone in men were largely unanticipated until recently, when oestrogen deficiency in males with aromatase deficiency and oestrogen resistance was found to cause osteoporosis and delayed fusion of epiphyses despite sufficient serum testosterone. This raises the possibility that in normal men oestrogens rather than androgens are of physiological importance in bone maturation. In the present study, we examined the association of serum oestradiol (E2) compared to that of free testosterone (FT) with bone mineral density (BMD) in normal men. The effect of oestrogen receptor (ER) gene polymorphism on BMD in men was also addressed.

SUBJECTS

Eighty‐one Thai men aged 20–79 years. All were healthy and did not take medication which may affect calcium and bone metabolism. BMD was assessed by DEXA. Dietary calcium was assessed by a 3‐day dietary record. Serum E2 and FT concentrations were measured by radioimmunoassay. Polymorphism at intron 1 of the α isoform of ER gene was determined by PCR–RFLP. Small p represents the presence of the restriction site while capital P indicates the absence of the restriction site.

RESULTS

Serum FT decreased with increasing age (r = −0.58, P < 0.0001) while E2 did not. However, there was a positive association between E2 and FT (r = 0.28, P < 0.05). Serum FT was related to BMD at femoral neck (r = 0.26, P < 0.05) and Ward's triangle (r = 0.30, P < 0.01) while E2 was related to BMD at anteroposterior (AP) lumbar spine (r = 0.29, P < 0.05), femoral neck (r = 0.23, P < 0.05) and femoral trochanter (r = 0.27, P < 0.05). Besides FT and E2, age, body weight, fat mass and fat‐free mass were also correlated to BMD at various skeletal sites. Using stepwise multiple linear regression to control for the confounding effects among these factors, fat‐free mass was found to be strongly associated with BMD at most skeletal sites. Serum E2 was related to BMD independently of other factors including FT at AP lumbar spine (r = 0.22, P < 0.05), femoral neck (r = 0.26, P < 0.01), femoral trochanter (r = 0.22, P < 0.05) and Ward's triangle (r = 0.26, P < 0.01) while serum FT was not associated with BMD at any site after controlling for E2 and other related factors. Concerning ERα gene polymorphism, 27 (33.3%) of the subjects had pp genotype, while 42 (51.9%) and 12 (14.8%) Pp and PP genotypes, respectively. After controlling for age, body weight, fat mass, fat‐free mass, calcium intake, FT and E2, the presence of P allele was associated with higher BMD at AP L2‐L4 (P < 0.05).

CONCLUSIONS

Serum oestradiol is more related to bone mass than free testosterone in normal men. Oestrogen‐receptor gene polymorphism is also associated with bone mass in men independently of oestradiol levels. Serum oestradiol together with oestrogen‐receptor genotype may partly determine bone mass in males.
Rosai‐Dorfman disease presenting as a pituitary tumour
Authors: Kelly W. F. , Bradey N. , Scoones D.
Published: March 2003

A 45‐year‐old woman had pyrexia, headaches, collapse and hyponatraemia. Intracerebral abscess, bacterial meningitis and subarachnoid haemorrhage were excluded. She was given intravenous antibiotics and gradually recovered. One month later she was readmitted with diplopia, headache and vomiting. Serum sodium was low (107 mmol/l) and a diagnosis of inappropriate ADH secretion was made. MRI scan showed a suprasellar tumour arising from the posterior pituitary gland. A skin rash gradually faded. Serum cortisol, prolactin, gonadotrophins and thyroid hormone levels were low. A pituitary tumour was removed trans‐sphenoidally, she had external pituitary radiotherapy, and replacement hydrocortisone and thyroxine. She was well for 12 months when she developed progressive weakness and numbness of both legs. Examination suggested spinal cord compression at the level of T2 where MRI scanning showed an intradural enhancing mass. This spinal tumour was removed and her neurological symptoms disappeared. Nine months after this she developed facial pain and nasal obstruction. CT scan showed tumour growth into the sphenoid sinus and nasal cavities. A right Cauldwell‐Luc operation was done and residual tumour in the nasal passages was treated by fractionated external radiotherapy and Prednisolone.

Histological examination of the specimens from pituitary, spinal mass, and nasal sinuses showed Rosai‐Dorfman disease, a rare entity characterized by histiocytic proliferation, emperipolesis (lymphophagocytosis) and lymphadenopathy. Aged 48 she developed cranial diabetes insipidus. Although Rosai–Dorfman syndrome is rare, it is being reported with increasing frequency, and should be borne in mind as a possible cause of a pituitary tumour.
Y‐chromosome identification by PCR and gonadal histopathology in Turner's syndrome without overt Y‐mosaicism
Authors: Mendes Judite R. T. , Strufaldi Maria Wany L. , Delcelo Rosana , Moisés Regina C. M. S. , Gilberto Vieira José , Kasamatsu Teresa S. , Francis Galera Marcial , Andrade Joyce A. D. , Verreschi Ieda T. N.
Published: March 2003

OBJECTIVE

The frequency of gonadoblastoma is high in patients with Turner's syndrome bearing cells with Y or partial Y‐chromosome. About 60% of patients with Turner's syndrome have a 45,X karyotype. In 30% of them a Y‐sequence is disclosed by DNA analysis. To identify patients at risk of developing gonadoblastoma, a PCR based assay with SRY, ZFY and DYZ3 specific primers was carried out to detect different Y‐sequences in the DNA of peripheral lymphocytes from patients with Turner's syndrome.

DESIGN AND PATIENTS

Peripheral blood karyotypes from 36 patients with Turner's syndrome were studied. Patients with proven Y‐chromosomal material were excluded. Genomic DNA was extracted from peripheral blood. SRY and ZFY genes and DYZ3 repetion of Y‐chromosome were amplified by PCR. Patients with clinical signs of hyperandrogenism or with positive Y‐sequences by PCR underwent gonadectomy. The gonadal tissues were examined for Y‐sequences using PCR, morphology and immunohistochemical study.

MEASUREMENTS

Turner's syndrome and signs of hyperandrogenism were evaluated both clinically and through laboratory tests. Haematoxylin and eosin staining was employed in gonadal morphology studies. The presence of testosterone was detected by immunohistochemistry using a monoclonal antibody.

RESULTS

Two patients who had Y‐positive blood samples and three hyperandrogenic (2 hirsutes, 1 virilized) Y‐negatives underwent gonadectomy. PCR was carried out on their gonadal tissue. The tissue from the two patients without hyperandrogenism was Y‐positive. The gonadal tissue from the three hyperandrogenics was Y‐negative. Gonadal morphology disclosed hilus cell hyperplasia in the 3 hyperandrogenic Y‐negatives and in one Y‐positive patient; stromal luteoma and hyperthecosis in the virilized patient, cystadenofibroma in one hirsute patient and gonadoblastoma in one Y‐positive. Testosterone was detected immunohistochemically in the hilus cell hyperplasia, stromal luteoma and hyperthecosis found in the hyperandrogenic patients.

CONCLUSIONS

The molecular study was sensitive and useful in the evaluation of patients at risk of developing gonadoblastoma. Other nontumour, gonadotrophin‐dependent and Y‐independent mechanisms which deserve the same medical approach may be involved in the genesis of hyperandrogenic signs in Turner's syndrome.