bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment of acute kidney allograft rejection with a non-mitogenic CD3 antibody
Authors: MEIJER R T , SURACHNO S , YONG S L , BEMELMAN F J , FLORQUIN S , TEN BERGE I J M , SCHELLEKENS P T A
Published: December 2004

T3/4.A is a non-mitogenic murine IgA mAb to human CD3 that was selected for clinical studies to provide an alternative for the mitogenic, T cell-activating, therapeutic mAb OKT3. Previously, we reported that T3/4.A is better tolerated in humans than the IgG2a-CD3 mAb T3/4.2a. Here we report the results of a phase II clinical trial to assess the immunosuppressive potential of T3/4.A. Eighteen first kidney transplant recipients with a first rejection episode were included. Baseline immunosuppression consisted of cyclosporin and prednisolone. Rejection treatment consisted of 5 mg mAb per day during 10 days. Fourteen patients responded, of whom four experienced a second rejection within 2 weeks, one experienced chronic rejection after 2·5 years, whereas the others remained rejection-free after treatment (median duration of follow-up 42 months). Four patients did not respond and eventually lost their graft. These results are similar to treatment results with OKT3, as reported in the literature. Following the first dose of T3/4.A, side effects were limited, and reduced compared to OKT3-treated controls. On the second day, 15 patients developed transient vomiting and/or diarrhoea, which coincided with elevated serum levels of proinflammatory cytokines. Minimal or even no side effects occurred during the remaining days, which is in sharp contrast to that seen generally during OKT3 treatment. Both T cell numbers and TCR expression were reduced during the therapy. We conclude that T3/4.A is a good alternative for OKT3 to treat rejection episodes in renal transplant recipients.
Increased numbers of CD38 molecules on bright CD8+ T lymphocytes in infectious mononucleosis caused by Epstein–Barr virus infection
Authors: ŽIDOVEC LEPEJ S , VINCE A , ÐAKOVIĆ RODE O , REMENAR A , JEREN T
Published: December 2004

The aim of this study was to quantify the expression of CD38 on CD8+ T lymphocytes of patients with infectious mononucleosis (IM) caused by Epstein–Barr virus (EBV) and cytomegalovirus (CMV). CD38 quantification technique chosen for this study was based on the enumeration of CD38 antibody binding sites in comparison to the quantification standards rather than determining relative fluorescence, which is difficult to standardize. The study enrolled 19 patients with typical clinical and laboratory parameters compatible with EBV-induced IM as well as 10 patients with atypical clinical presentation of this disease. Furthermore, CD38 expression was analysed in a group of 13 patients with IM caused by CMV infection. CD38 quantification was performed within 6 days of the presentation of symptoms. All three groups of IM patients showed a statistically significant increase in the number of anti-CD38 antibody binding sites (which correspond to the number of CD38 molecules) on bright CD8+ T lymphocytes compared to healthy controls. The numbers of CD38 molecules expressed on CD8+ T lymphocytes did not differ significantly between IM patients with typical and atypical clinical presentation of the disease. Patients with CMV-induced IM had significantly lower numbers of CD38 molecules expressed on CD8+ T lymphocytes. Therefore, we conclude that CD38 quantification could be helpful in differential diagnostics of IM cases with atypical clinical presentation.
Hypogalactosylation of serum IgG in patients with coeliac disease
Authors: CREMATA J A , SORELL L , MONTESINO R , GARCÍA R , MATA M , CABRERA G , GALVAN J A , GARCÍA G , VALDÉS R , GARROTE J A
Published: December 2004

Coeliac disease (CD) is described as an autoimmune enteropathy associated with the presence of IgG and IgA antigliadin and antitransglutaminase autoantibodies. While of diagnostic significance, the role of these autoantibodies in the immunopathogenesis of CD is elucidated. An inappropriate T cell immune response to gluten is also involved in the pathogenesis of CD, as evidenced by autoantibody switching. The N-glycans released from serum IgG of CD patients and three groups of healthy controls, of differing age ranges, were analysed by NH2-high performance liquid chromatography (HPLC). The fucosylated biantennary N- glycans were the most abundant neutral oligosaccharides; in particular, the agalacto form (G0F) showed a mean value of 42% (s.d. ± 7·4), 30% (s.d. ± 5·9), 26% (s.d. ± 4·2) and 35% (s.d. ± 6·8) for CD patients, healthy children, healthy adults under 40 and healthy adults over 40 years old, respectively. The ratio of asialo agalacto fucosylated biantenna to asialo monogalacto fucosylated biantenna (G0F)/(G1F) for CD patients showed a significant increase compared to healthy children (P < 0·0002), healthy adults under 40 (P < 0·0002) and healthy adults over 40 years old (P < 0·01). Hypogalactosylation was more pronounced for CD patients than for the patients with other autoimmune diseases such as rheumatoid arthritis or psoriatic arthritis.
Critical roles of CD30/CD30L interactions in murine autoimmune diabetes
Authors: CHAKRABARTY S , NAGATA M , YASUDA H , WEN L , NAKAYAMA M , CHOWDHURY S A , YAMADA K , JIN Z , KOTANI R , MORIYAMA H , SHIMOZATO O , YAGITA H , YOKONO K
Published: December 2004

CD30/CD30L is a member of tumour necrosis factor (TNF) receptor/TNF superfamily and has been implicated in immune-regulation. A genetic study has also suggested a possible implication of CD30 in spontaneous autoimmune diabetes in NOD mice. In this study, we investigated the involvement of CD30/CD30L in the development of diabetes in NOD mice. Flow cytometric analysis showed that CD30 and CD30L were highly expressed on CD4+ or CD8+ T cells in the spleen and pancreatic lymph node of younger NOD mice. In addition, islet-specific CD4+ or CD8+ T cell lines expressed CD30 and CD30L. Administration of a neutralizing anti-CD30L monoclonal antibody (mAb) from 2 to 10 week of age completely suppressed the development of spontaneous diabetes in NOD mice. In addition, the treatment with anti-CD30L mAb also inhibited the development of diabetes induced by adoptive transfer of spleen cells from diabetic NOD mice or islet-specific CD4+ or CD8+ T cell lines into NOD-SCID mice. Furthermore, anti-CD30L mAb inhibited T cell proliferation in response to islet antigens. These results suggested that CD30/CD30L interaction plays important roles in both induction and effector phases of autoimmune diabetes in NOD mice.
Predictive markers for development of strongyloidiasis in patients infected with both Strongyloides stercoralis and HTLV-1
Authors: SATOH M , KIYUNA S , SHIROMA Y , TOMA H , KOKAZE A , SATO Y
Published: December 2004

Severe strongyloidiasis has often been reported to occur in some patients infected with both Strongyloides stercoralis (S. stercoralis) and human T-cell leukaemia virus type 1 (HTLV-1); however, there are few useful predictive markers for the risk of development of strongyloidiasis in these patients. To search for such predictive markers, we examined peripheral blood and stool samples of individuals infected with both S. stercoralis and HTLV-1 in Okinawa, Japan, an area in which both of these are endemic. The HTLV-1 proviral load and antibody titre were examined in relation to the S. stercoralis load as measured by the direct faecal smear method in patients infected with both S. stercoralis and HTLV-1. The Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) antibody titre was also measured in these patients in order to examine the relationship between host immunity and HTLV-1 proviral load or antibody titre. The direct faecal smear-positive group showed both a higher HTLV-1 proviral load and HTLV-1 antibody titre than the -negative group (P < 0·05). In contrast, inverse correlations of these parameters with the EBNA antibody titre were observed, especially for proviral load (ρ = −0·387, P <  0·05). These results suggest that HTLV-1 proviral load and antibody titre influence the S. stercoralis load via disturbance of the host immunity, and that proviral load would be an especially useful predictive marker of the risk of development of strongyloidiasis in patients infected with both S. stercoralis and HTLV-1.
CD8β/CD28 expression defines functionally distinct populations of peripheral blood T lymphocytes
Authors: WERWITZKE S , TIEDE A , DRESCHER B E , SCHMIDT R E , WITTE T
Published: December 2004

Peripheral blood CD8+ T lymphocytes generally express the CD8 coreceptor as an αβ heterodimer. On these cells, the CD8β chain is present either at high (CD8βhigh) or low density (CD8βlow). CD8βhigh cells are CD28+, whereas CD8βlow cells are CD28+ or CD28–. Therefore, three subpopulations of CD8+ T cells can be described: (i) CD8βhighCD28+ (ii) CD8βlowCD28+, and (iii) CD8βlowCD28– cells. Phenotypic and functional characterization of these CD8+ T cell subsets revealed significant differences. CD8βhighCD28+ cells predominantly express CD45RA. In contrast, CD8βlowCD28+ cells frequently express CD45R0 and the activating NK receptor CD161. CD8βlowCD28– cells frequently revert to the CD45RA phenotype. In addition, these cells express CD16, CD56, CD94, and the killer-inhibitory receptors NKB1 and CD158a. Intracellular IL-2 was frequently detected in CD8βhighCD28+ cells and CD8βlowCD28+ cells, but not CD8βlowCD28– cells. CD8βlowCD28+ cells and CD8βlowCD28– cells frequently stained positive for IFN-γ. In addition, these cells contain intracellular perforin and granzyme A. Expression of Fas (CD95) as well as susceptibility to apoptosis is markedly increased in CD8βlowCD28+ and CD8βlowCD28– cells as compared to CD8βhighCD28+ cells. In vitro activation of peripheral blood lymphocytes triggered expansion of CD8βhighCD28+ cells as well as a development into CD8βlowCD28+ and CD8βlowCD28– cells. Similarly, activation of CD8βhighCD28+ cord blood cells resulted in the appearance of CD8βlowCD28+ and CD8βlowCD28– cells. These data suggest that CD8βhighCD28+ cells can differentiate into CD8βlowCD28+ and CD8βlowCD28– cells upon TCR stimulation. Therefore, the CD8β/CD28 subsets in peripheral blood may reflect distinct stages of post-thymic CD8+T cell development.
Mannan-binding lectin in children with Escherichia coli O157:H7 haemmorrhagic colitis and haemolytic uraemic syndrome
Authors: PROULX F , WAGNER E , TOLEDANO B , DECALUWE H , SEIDMAN E G , RIVARD G-E
Published: December 2004

Mannan-binding lectin (MBL) triggers complement activation upon binding to microbial surfaces. MBL deficiency has been associated with increased susceptibility to severe bacterial infections. We hypothesized that MBL deficiency may predispose children to Shiga toxin-producing Escherichia coli (STEC) O157:H7 infections and the associated haemolytic uraemic syndrome (HUS). We compared circulating levels of MBL among children with uncomplicated O157:H7 haemorrhagic colitis (HC), patients with O157:H7 HUS, normal and diseases control groups. Circulating MBL concentrations on admission were as follows: 3·22 ± 2·43 µg/ml among normal controls (n = 23); 2·90 ± 2·44 µg/ml in patients with rotavirus enteritis (n = 10); 2·78 ± 1·65 µg/ml in children with HC due to non-STEC bacterial pathogen (n = 15); 2·67 ± 2·44 µg/ml in patients with uncomplicated O157:H7 HC (n = 27); 2·80 ± 2·97 µg/ml in children with O157:H7 HUS (n = 15); 6·70 ± 4·49 µg/ml in patients with chronic renal failure unrelated to O157:H7 infection (n = 6). Higher MBL levels were found in patients with chronic renal failure compared to O157:H7 HC (P < 0·047). However, MBL concentrations <0·5 µg/ml, which have been associated with MBL deficiency in relation to increased susceptibility to infections, were noted at comparable rates between the different groups (P = NS). Our data does not support that MBL deficiency may predispose to O157:H7 infections nor than the development of diarrhoea associated HUS.
Measurement and interpretation of pneumococcal IgG levels for clinical management
Authors: BALMER P , NORTH J , BAXTER D , STANFORD E , MELEGARO A , KACZMARSKI E B , MILLER E , BORROW R
Published: December 2004

The detection of pneumococcal IgG antibodies is helpful for the evaluation of response to pneumococcal vaccination and need for revaccination. Results generated by the clinical assay which is currently used, in which the 23 valent polysaccharide vaccine is the antigen, were compared to those obtained by a capsular polysaccharide serotype-specific assay that measures IgG antibodies to 9 common serotypes causing invasive disease. Discrepancies in 21/47 (45%) of the results were observed in a direct comparison between the two assays. In each case a positive titre was obtained on the clinical assay but IgG levels on the serotype-specific assay were below the putative protective level of 0·2 µg/ml for at least one of the 9 serotypes assayed. The generation of false positives by the current clinical assay is due to its lack of specificity. Antibodies to C-polysaccharide and all of the 23 serotypes included in the pneumococcal polysaccharide vaccine are incorporated into the final titre whereas the serotype-specific assay adsorbs out noncapsular polysaccharide antibodies. The discrepancies between the two assays highlight the importance of standardized assays that measure putative correlates of protection and demonstrate the need to re-evaluate the current clinical assay. A tool that allows the interpretation of the results of the serotype-specific assay is provided and its potential for assessing individual susceptibility levels to vaccine preventable pneumococcal infection is discussed.
Humoral β-cell autoimmunity is rare in patients with the congenital rubella syndrome
Authors: VISKARI H , PARONEN J , KESKINEN P , SIMELL S , ZAWILINSKA B , ZGORNIAK-NOWOSIELSKA I , KORHONEN S , ILONEN J , SIMELL O , HAAPALA A-M , KNIP M , HYÖTY H
Published: December 2004

The congenital rubella syndrome (CRS) is associated with increased risk for diabetes and thyroid disease. However, the mechanisms by which the rubella virus may cause these diseases are poorly characterized. Previous studies were carried out before modern immunological methods were available. The present study aimed at evaluating whether autoimmune mechanisms are involved in the pathogenesis by analysing antibodies to biochemically characterized autoantigens. The incidence of clinical diabetes, thyroid disease, coeliac disease and related antibodies (islet cell antibodies, ICA; insulin autoantibodies, IAA; antibodies to the tyrosine phosphatase related IA-2 molecule, IA-2 A and glutamic acid decarboxylase, GADA; thyroid peroxidase, TPO; tissue transglutaminase, TTGA; and gliadin, AGA) and HLA risk genotypes were analysed in 37 subjects affected by or exposed to rubella during fetal life (mean age 22·5 years). One patient had diabetes and four patients had clinical hypothyroidism at the time of the examination. ICA, IAA, GADA or IA-2 A were not detected in any of the patients, while five patients tested positive for TPO antibodies. Coeliac disease or TTGA were not observed. Eight patients carried the HLA-DR3–associated HLA-DQB1*02-DQA1*05 haplotype. These results provide no evidence of an increased frequency of markers for humoral β-cell autoimmunity in patients with CRS suggesting that diabetes in CRS may be caused by other than autoimmune mechanisms.
Donor dependent, interferon-γinduced HLA-DR expression on human neutrophilsin vivo
Authors: REINISCH W , LICHTENBERGER C , STEGER G , TILLINGER W , SCHEINER O , GANGL A , MAURER D , WILLHEIM M
Published: December 2004

Neutrophils are effector cells of innate immune responses. Stimulated by interferon-γ (IFN-γ) to express HLA-DR, neutrophils acquire accessory cell functions for superantigen-mediated T cell activation. In vitro HLA-DR induction on neutrophils varies in a functionally relevant way as levels of MHC class II expression and magnitude of neutrophil induced T cell responses are correlated functions. The aim of this study was to assess whether IFN-γ induces HLA-DR on human neutrophils in a donor dependent fashion in vivo and to define regulatory events operative in MHC class II expression of neutrophils. In vivo administration of rhIFN-γ in 55 patients with renal cell carcinoma resulted in a varying increase of HLA-DR on neutrophils. By setting a cut-off for response at>10% HLA-DR positive neutrophils, HLA-DR responders (51%) were as frequent as nonresponders (49%). In vivo kinetic studies revealed a peak expression of HLA-DR on neutrophils 48 h after rhIFN-γ application, while nonresponders remained HLA-DR negative over a 72-h period. In vitro IFN-γ stimulated neutrophils recapitulated the response profiles observed in vivo. No differences in IFN-γ dependent CD64 and invariant chain expression, and IFN-γ serum levels were observed among the response subgroups. HLA-DR mRNA was detected in neutrophils from rhIFN-γ treated responders and nonresponders, HLA-DR protein solely in lysates of responder neutrophils. IFN-γ stimulated HLA-DR expression on neutrophils is subject to donor dependent variations in vivo, which result from rather post-transcriptional than transcriptional regulation. Due to their abundance in inflammatory reactions heterogeneous HLA-DR expression by neutrophils could determine the outcome of superantigen-driven diseases.
Abnormalities in the T and NK lymphocyte phenotype in patients with Nijmegen breakage syndrome
Authors: MICHAŁKIEWICZ J , BARTH C , CHRZANOWSKA K , GREGOREK H , SYCZEWSKA M , WEEMAES C M B , MADALIŃSKI K , DZIERŻANOWSKA D , STACHOWSKI J
Published: December 2004

Nijmegen breakage syndrome (NBS) is a rare autosomal recessive disorder characterized by spontaneous chromosomal instability with predisposition to immunodeficiency and cancer. In order to assess the cellular basis of the compromised immune response of NBS patients, the distribution of functionally distinct lymphocyte subsets in peripheral blood was evaluated by means of double-colour flow cytometry. The study involved the 36 lymphopenic patients with a total lymphocyte count ≤1500 µl (group A) and seven patients (group B) having the absolute lymphocyte count comparable with the age-matched controls (≥3000 µl). Regardless of the total lymphocyte count the NBS patients showed: (1) profound deficiency of CD4+ and CD3/CD8+ T cell subsets and up to fourfold increase in natural killer (NK) cells, almost lack of naive CD4+ T cells expressing CD45RA isoform, unchanged percentage of naive CD8+ cell subset (CD8/CD45RA+) but bearing the CD8 receptor of low density (CD8low); (2) normal expression of CD45RA isoform in the CD56+ lymphocyte subset, profound decrease in αβ but up to threefold increase in γδ-T cell-receptor (TCR)-positive T cells; (3) shift towards the memory phenotype in both CD4+ and CD8+ lymphocyte subpopulations expressing CD45RO isoform (over-expression of CD45RO in terms of both the fluorescence intensity for CD45RO isoform and the number of positive cells); and (4) an increase in fluorescence intensity for the CD45RA isoform in NK cells population. These results indicate either a failure in T cell regeneration in the thymic pathway (deficiency of naive CD4+ cells) and/or more dominant contribution of non-thymic pathways in lymphocyte renewal reflected by an increase in the population of CD4+ and CD8+ memory cells, γδ-TCR positive T as well as NK cell subsets.
Bruton's tyrosine kinase defect in dendritic cells from X-linked agammaglobulinaemia patients does not influence their differentiation, maturation and antigen-presenting cell function
Authors: GAGLIARDI M C , FINOCCHI A , ORLANDI P , CURSI L , CANCRINI C , MOSCHESE V , MIYAWAKI T , ROSSI P
Published: December 2004

X-linked agammaglobulinaemia (XLA) is a primary immunodeficiency disease characterized by very low levels or even absence of circulating antibodies. The immunological defect is caused by deletions or mutations of Bruton's tyrosine kinase gene (Btk), whose product is critically involved in the maturation of pre-B lymphocytes into mature B cells. Btk is expressed not only in B lymphocytes but also in cells of the myeloid lineage, including dendritic cells (DC). These cells are professional antigen presenting cells (APC) that play a fundamental role in the induction and regulation of T-cell responses. In this study, we analysed differentiation, maturation, and antigen-presenting function of DC derived from XLA patients (XLA-DC) as compared to DC from age-matched healthy subjects (healthy-DC). We found that XLA-DC normally differentiate from monocyte precursors and mature in response to lipopolysaccharide (LPS) as assessed by de novo expression of CD83, up-regulation of MHC class II, B7·1 and B7·2 molecules as well as interleukin (IL)-12 and IL-10 production. In addition, we demonstrated that LPS stimulated XLA-DC acquire the ability to prime naïve T cells and to polarize them toward a Th1 phenotype, as observed in DC from healthy donors stimulated in the same conditions. In conclusion, these data indicate that Btk defect is not involved in DC differentiation and maturation, and that XLA-DC can act as fully competent antigen presenting cells in T cell-mediated immune responses.
The diagnostic use of the rhTSH/thyroglobulin test in differentiated thyroid cancer patients with persistent disease and low thyroglobulin levels
Authors: Pellegriti Gabriella , Scollo Claudia , Regalbuto Concetto , Attard Marco , Marozzi Paola , Vermiglio Francesco , Violi Maria Antonella , Cianci Michelangela , Vigneri Riccardo , Pezzino Vincenzo , Squatrito Sebastiano
Published: December 2004

background Serum thyroglobulin (Tg) measurement after TSH stimulation, by either thyroid hormone withdrawal or recombinant human TSH (rhTSH) administration, is the most sensitive method for early detection of patients with persistent or recurrent differentiated thyroid cancer (DTC) after total thyroidectomy and 131I ablation. The use of rhTSH is now increasing because it avoids thyroid hormone suppressive therapy (THST) withdrawal and the consequent symptoms of severe hypothyroidism. Current guidelines suggest measurement of serum Tg 4 days after starting a 2‐day course of rhTSH injections, and assumes that Tg reaches maximum serum levels at that time.

objective The present study was carried out to evaluate the accuracy of rhTSH/thyroglobulin test in DTC patients with persistent disease and low thyroglobulin levels.

patients and measurements A series of 13 DTC patients was selected because they had proven persistent disease associated with low Tg levels (< 2·0 µg/l) under l‐thyroxine treatment. In all of them, serum Tg was > 5·0 µg/l at the last THST withdrawal. We measured serum Tg and TSH levels on days 0·5, 1, 1·5, 2, 4, 7, 10 and 15 after the first of a 2‐day course of intramuscular rhTSH injections.

results Serum Tg values were variable in terms of both peak and time‐course. Detectable serum Tg levels were recorded on day 4 in all patients. However, among these 13 patients, the peak Tg value was reached earlier than day 4 in three patients and later in two others. In one patient, Tg level at day 2 was higher (3·0 µg/l) than at day 4 (1·8 µg/l). In six of the 13 patients studied we compared Tg values after rhTSH to those subsequently obtained after THST withdrawal: in five of them Tg values were two to three times higher after the latter stimulation. Serum Tg value variability after rhTSH was partially accounted for by variability of serum TSH levels, which were inversely related to patient body surface.

conclusions In DTC patients with persistent disease and low Tg levels, optimization of the diagnostic use of Tg measurement after rhTSH may require rhTSH dose adjustment to the patient body surface area and repeated blood sampling, in order to improve diagnostic accuracy. In these patients not even a TSH‐stimulated serum Tg cut‐off of 2·0 µg/l on day 4 provides 100% accuracy, whereas a cut‐off of 1·0 µg/l seems more appropriate. Therefore, in this subset of patients, if any detectable Tg level ≥ 1·0 µg/l is found after rhTSH, re‐evaluation after THST should be advised.
Low cholesteryl ester transfer protein (CETP) concentration but normal CETP activity in serum from patients with short‐term hypothyroidism Lack of relationship to lipoprotein abnormalities
Authors: Dedecjus Marek , Masson David , Gautier Thomas , De Barros Jean‐Paul Pais , Gambert Philippe , Lewinski Andrzej , Adamczewski Zbigniew , Moulin Philippe , Lagrost Laurent
Published: December 2004

objectives Hypothyroidism is associated with a number of abnormalities in lipoprotein metabolism. Although alterations in neutral lipid exchanges among plasma lipoproteins might be one characteristic feature of hypothyroidism, a few human studies of cholesteryl ester transfer protein (CETP) activity have led to heterogeneous and fragmentary observations. The aim of the present study was to analyse the influence of short‐term hypothyroidism on CETP activity, as well as on the structure and composition of lipoproteins.

patients, design and measurements Sixty‐six thyroidectomized patients were withdrawn from L‐thyroxine (L‐T4) treatment for 5 weeks. Subsequently, L‐T4 therapy was reinstated for 2 months and patients were compared to 61 matched normolipidaemic controls. Serum CETP activity and mass concentration, serum lipids, apolipoproteins and lipoprotein size distribution were determined in the three groups.

results Serum CETP mass concentration was significantly decreased in short‐term hypothyroid patients, as compared to control subjects (3·22 ± 0·98 vs. 3·79 ± 1·2 mg/l, respectively; P < 0·001), and the values were normalized during L‐T4 therapy. The ability of endogenous serum lipoproteins to interact with CETP was normal in short‐term hypothyroid patients. Concordant observations were made regardless of whether neutral lipid transfers were measured from high‐density lipoproteins (HDL) toward apo B‐containing lipoproteins or from liposomes toward HDL. The size distribution of HDL was significantly different in short‐term hypothyroid patients, compared to either the control or treated subgroups, with significant higher proportions of large‐sized HDL2b and HDL2a (HDL2b: 13·6 ± 6·5% before vs. 8·5 ± 4·2% during L‐T4 therapy, P < 0·05; HDL2a, 33·0 ± 7·0% before vs. 29·3 ± 6·9% during L‐T4 therapy, P < 0·05). Although serum CETP mass concentration correlated negatively with the HDL2 to HDL3 ratio in control subjects (r = −0·588; P < 0·0001), no significant correlations were observed in hypothyroid patients, regardless of whether they were treated or not. Similarly, whereas the previously recognized positive correlation of CETP mass concentration with serum LDL cholesterol levels was found in control subjects (r = 0·264; P < 0·05), no significant correlations appeared in treated and untreated patients.

conclusions Short‐term hypothyroidism may constitute an unique situation in which concomitant alterations in serum cholesteryl ester transfer protein concentration and lipoprotein parameters are disconnected.
Blood pressure in children and adolescents with congenital adrenal hyperplasia (21‐hydroxylase deficiency): a preliminary report
Authors: Roche Edna F. , Charmandari Evangelia , Dattani Mehul T. , Hindmarsh Peter C.
Published: December 2004

background Classical congenital adrenal hyperplasia (CAH) is characterized by a defect in cortisol and aldosterone secretion, adrenal hyperandrogenism, impaired adrenal medullary function and insulin insensitivity. The latter along with the increased tendency towards obesity raises questions whether other cardiovascular risk factors are altered in CAH.

objective To evaluate 24‐h ambulatory blood pressure and obesity in patients with salt‐wasting 21‐hydroxylase deficiency diagnosed in the neonatal period and treated with hydrocortisone and 9α‐fludrocortisone thereafter.

methods Thirty‐eight children (15 males) aged 11·2 years (range 6·1–18·2 years) underwent 24‐h ambulatory blood pressure monitoring in the hospital setting. Standard anthropometric measures of height, weight and skinfold thickness were undertaken and body mass index (BMI) derived. All data were expressed as standard deviation scores (SDS) using the UK Growth Reference data.

results Mean daytime systolic blood pressure SDS (1·8, SD 1·1) was significantly higher than the reference population (P < 0·001), and 58% of patients (67% males; 52% females) had systolic hypertension. Mean daytime diastolic blood pressure SDS (0·8, SD 0·8) was also elevated and 24% (13% males; 37% females) had diastolic hypertension. Eighty‐four per cent had absence of the physiological nocturnal dip in systolic blood pressure. Height SDS was similar to the reference population but BMI SDS was higher (P < 0·001). BMI SDS was related to systolic blood pressure SDS (r = 0·34; P = 0·03) and the effect was most marked in females where it was related to measures of truncal fat (r = 0·82; P = 0·002).

conclusions Children with salt‐wasting 21‐hydroxylase deficiency have elevated 24‐h ambulatory blood pressure and absence of the physiological nocturnal dip in blood pressure. These abnormalities are associated with a raised BMI, particularly in females. Regular measurement and plotting of blood pressure should be part of the management of children with classical CAH.
Multiple endocrine neoplasia type 1 (MEN1) germline mutations in familial isolated primary hyperparathyroidism
Authors: Pannett A. A. J. , Kennedy A. M. , Turner J. J. O. , Forbes S. A. , Cavaco B. M. , Bassett J. H. D. , Cianferotti L. , Harding B. , Shine B. , Flinter F. , Maidment C. G. H. , Trembath R. , Thakker R. V.
Published: December 2004

background Familial isolated hyperparathyroidism (FIHP) is an autosomal dominant disorder characterized by uniglandular or multiglandular parathyroid tumours that occur in the absence of other endocrine tumours. The disorder may represent either an early stage of multiple endocrine neoplasia type 1 (MEN1), or an allelic variant of MEN1, or a distinct entity involving another locus. We have explored these possibilities in seven families in whom primary hyperparathyroidism occurred as the sole endocrinopathy.

methods Seven FIHP families were ascertained and venous blood samples obtained from 35 members (17 affected and 18 unaffected) for DNA sequence analysis of the MEN1 gene. The mean (± SD) follow‐up period in the 17 affected members was 15·06 (± 8·83) years.

results Four heterozygous germline mutations of the MEN1 gene were identified. These consisted of two 4‐bp intragenic deletions that would result in prematurely truncated proteins, and two missense (Asp153Val and Ala411Pro) mutations. Furthermore, analysis of parathyroid tumour DNA from one individual revealed a loss of the wild‐type allele and retention of the mutant allele, consistent with Knudson's ‘two‐hit’ model of hereditary cancer and a tumour suppressor role for MEN1 in FIHP.

conclusions Our results provide further support for FIHP being a distinct allelic variant of MEN1, and an analysis of the 16 mutations reported to date indicate that FIHP is associated with a higher frequency of missense MEN1 mutations.
Evaluation of the reproducibility of the GHRH plus GHRP‐6 test of growth hormone reserve in adults
Authors: Popovic Vera , Pekic Sandra , Micic Dragan , Damjanovic Svetozar , Marikovic Jelena , Simic Mirjana , Dieguez Carlos , Casanueva Felipe F.
Published: December 2004

background  The diagnosis of GH deficiency (GHD) is controversial, relying on GH secretion elicited by the provocative tests of GH reserve. However, the performance of most of the tests in use have not been evaluated rigorously. The repeatability of a test is a prerequisite before evaluating its diagnostic capability.

objective  The combined administration of GH‐releasing hormone (GHRH) and GH‐releasing hexapeptide (GHRP‐6) is an efficacious test of GH reserve in adults, and the target of this work was to evaluate its reproducibility.

methods  Seventeen healthy subjects were challenged with GHRH plus GHRP‐6 (1 µg/kg i.v.). All subjects underwent four tests on different days separated by at least 2 months. GH peaks were evaluated by several mathematical analyses of reproducibility.

results  As a group, the subjects showed high reproducibility after the four tests, with GH peaks of 46·0 ± 5·1; 48·4 ± 6·4; 50·1 ± 5·4 and 52·9 ± 5·8 µg/l, respectively (1 µg/l = 3 mU/l). Individually analysed, the reproducibility was good, and the regression analysis showed a correlation between tests 1 and 2 of R = 0·729, P < 0·0009, between tests 1 and 3 of R = 0·710, P < 0·001, and between tests 1 and 4 of R = 0·683, P < 0·002. Under mathematical analysis, the multiple correlation coefficient analysis, analysis of variance (anova) with repeated measurements, repeatability index, the simplest coefficient of variation and the intraclass correlation coefficient (ICC) all unambiguously showed that the GHRH + GHRP‐6 test was reproducible. Furthermore, the repeated tests did not alter the biochemical diagnosis of the subjects, with absence of false‐positive values.

conclusions  The GHRH + GHRP‐6 test of GH reserve is reproducible in adult subjects.
Oestrogen receptor α genotype, and interactions between vitamin D receptor and transforming growth factor‐β1 genotypes are associated with quantitative calcaneal ultrasound in postmenopausal women
Authors: Koh Jung‐Min , Nam‐Goong Il Seong , Hong Jeong Soo , Kim Hong‐Kyu , Kim Jae Seung , Kim Shin‐Yoon , Kim Ghi Su
Published: December 2004

objective  Quantitative ultrasound (QUS) of bone is a new radiation‐free, low‐cost method that measures both bone mass and quality. We investigated associations between QUS parameters and polymorphisms of vitamin D receptor (VDR), oestrogen receptor α (ERα) and transforming growth factor‐β1 (TGF‐β1) genes in postmenopausal women residing in a community.

design  QUS and anthropometric characteristics were measured in postmenopausal women, and compared with regard to the VDR, ERα and TGF‐β1 genotypes.

patients  Among the 552 women who participated in the population‐based Chung‐Up osteoporosis prevalence study, 206 postmenopausal women, aged 60–69 years, were included.

measurements  Broadband ultrasound attenuation (BUA) and speed of sound (SOS) were measured at the left calcaneus using QUS measurement of bone, and a stiffness index (SI) was calculated. We determined the BsmI and FokI polymorphisms of VDR gene and the XbaI and PvuII polymorphisms of ERα gene using the polymerase chain reaction‐restriction fragment length polymorphism method, and Τ29 → C polymorphism of TGF‐β1 gene using an allele‐specific polymerase chain reaction assay.

results  The XbaI polymorphism of ERα gene was significantly associated with SI (T‐score) and BUA (P = 0·040 and P = 0·024, respectively). There were no significant differences in any QUS parameters among the genotypes of VDR and TGF‐β1. However, significant genetic interactions between the VDR and TGF‐β1 genotypes, were noted (P = 0·017 for SI and P = 0·028 for BUA between the BsmI and Τ29 → C polymorphisms; P = 0·038 for SI and P = 0·035 for BUA between the FokI and T29 → C polymorphisms). The combined genotypes between the BsmI and T29 → C polymorphisms or between the FokI and T29 → C polymorphisms, were significantly associated with the QUS parameters.

conclusions  This study indicates that the XbaI polymorphism of ERα gene may influence the Quantitative ultrasound parameters in postmenopausal women, and suggests the need for further investigations about the interactions between the VDR and TGF‐β1 genes.
Assessment of GH reserve before and after successful treatment of adult patients with Cushing's syndrome
Authors: Tzanela Marinella , Karavitaki Niki , Stylianidou Christiana , Tsagarakis Stylianos , Thalassinos Nikos C.
Published: December 2004

objective  Several studies have demonstrated impaired GH secretion in patients with active Cushing's syndrome (CS). It has been suggested that persistence of GH deficiency, despite treatment of cortisol excess, may delay the recovery of these patients and therefore temporary treatment with GH may have some benefit. However, the time course of restoration of GH secretion after successful treatment of CS has only been investigated in a limited number of mostly paediatric reports. The aim of the present study was the evaluation of GH reserve in adult patients with CS before and after correction of cortisol excess.

design and patients  Sixteen patients (12 females, four males) with CS aged 44·7 ± 5·05 years were recruited. These included seven patients with Cushing's disease, four patients with ectopic ACTH secretion and five patients with adrenal adenoma. All patients were evaluated before any therapeutic intervention. Twelve patients were successfully treated following appropriate surgery and these were further studied. The combined pyridostigmine/GHRH test was used to assess GH reserve in these patients. In a proportion of cases an insulin tolerance test (ITT) was also used.

results  Before any therapeutic intervention, an impaired GH response to PD/GHRH was noted in all patients. Restoration of GH response at 6 months was observed in six patients (50%); at 12 months in two; at 18 months in one patient. Two of the patients with no restoration of GH response at 12 months did not accept further investigation. Only one patient did not achieve an adequate GH response even when tested 30 months following cure of CS. Restoration of GH reserve was more commonly observed in those patients in whom there was recovery of the HPA axis. There was a good correlation between peak GH levels to PD + GHRH and ITT. No statistically significant difference was revealed in IGF‐I levels between pre‐ and post‐treatment evaluation.

conclusions  Adult patients with active Cushing's syndrome demonstrate a profound suppression of stimulated GH secretion. In the majority of these patients the disruption of GH secretion is normalized within a year after successful treatment of endogenous cortisol excess.
Video fluorescein imaging of the skin: description of an overviewing technique for functional evaluation of regional cutaneous blood perfusion in occlusive arterial disease of the limbs
Authors: Lund F. , Jogestrand T. , Inacio J. , Larsson U. B. , Lund S. , Sörhed K.
Published: December 2004

In peripheral arterial occlusive disease (PAOD), and particularly in critical limb ischaemia, fluorescence recording of the skin after i.v. injection of sodium fluorescein provides an overviewing functional imaging of regional blood perfusion as revealed by a continuous, almost geographical, mapping of changes in the dynamic fluorescence pattern. Based on the previously used technique of rapid sequence flash photography introduced by Lund and Lund in 1973b, a new video fluorescence imaging (VFI) technique has been developed and used in clinical routine. VFI has a broad versatility with non‐touch access to areas of different size, such as the feet, hands or larger limb areas. Examples of application are the diagnosis of small artery disease, predictive prognosis of critical limb ischaemia, evaluation of distal outflow in connection with arterial reconstruction or pharmacotherapy of non‐reconstructable critical ischaemia, as well as in decisions on amputation and assessment of amputation level. Thanks to its overviewing qualities with reproducible mapping of blood perfusion in standardized examination conditions, VFI would seem to have a well‐justified role in angiology and above all in PAOD in both research and clinical routine work.
Reproducibility of a semiautomated acetylene rebreathing technique for measuring cardiac output in humans at rest
Authors: Hunt B. E. , Davy K. P. , Seals D. R.
Published: December 2004

The specific aims of the present study were to determine: (1) the day‐to‐day reproducibility of a semiautomated acetylene rebreathing technique for measuring cardiac output under resting conditions; (2) the reproducibility of this technique among subjects differing in gender and age; and (3) the number of trials within a session necessary to maximize the day‐to‐day reproducibility of the technique. To address these aims, cardiac output was measured in 21 healthy men (n = 8) and women (n = 13) between the ages of 25 and 71 years in the supine posture on two separate days. Mean levels of cardiac output at rest were similar on day 1 vs. day 2 in the overall group. Cardiac output measured on day 1 was highly correlated (r = 0·98, P<0·001) with cardiac output measured on day 2. The day 1 to day 2 mean difference in cardiac output for the individual subjects was <4%. The mean levels of heart rate and stroke volume also were similar between day 1 and day 2. The relation between cardiac output measured on day 1 vs. day 2 in the gender and age subgroups was similar to that observed in the overall group. The mean absolute difference among the three rebreathing trials within a day was 360 ml min−1 in the overall group, with a coefficient of variation of 7%. The variability between rebreathing trials measured on day 1 vs. day 2 in the gender and age subgroups was similar to that observed in the overall group. The reliability of cardiac output measured on different days was excellent with a single rebreathing trial (r = 0.93) and improved significantly up to three trials (r = 0.98). In conclusion, the findings of the present study indicate that the acetylene rebreathing technique can be a highly reproducible method for measuring cardiac output under resting conditions. The reproducibility is consistently strong in healthy humans of varying age and in both genders, and is enhanced by the use of multiple trials.
Scintigraphic assessment of focal platelet accumulations following infrainguinal bypass surgery in humans
Authors: Nielsen T. G. , Hesse B. , Eiberg J. , Rabøl A. , Folke M. , Schroeder T. V.
Published: December 2004

Enothelial injury is assumed to be of pathogenetic significance in the development of graft stenoses, which remain a major cause of failure of peripheral bypasses. The aim of this study was to assess endothelial injury related to infrainguinal bypass surgery by indium‐111 platelet scintigraphy. In 28 patients undergoing in situ vein (n = 24), composite vein‐polytetrafluoroethylene (PTFE) (n = 1) or PTFE (n = 3) bypass surgery, assumed vascular injuries were recorded intraoperatively. Autologous indium‐111‐labelled platelets were injected into the inflow artery immediately after restoration of flow in the graft. Platelet deposition was assessed by gamma‐camera images of thigh and crus obtained 4 and/or 24 h after surgery. Areas of focally increased activity were recorded and graded as moderate or intense. In the 24 vein bypasses, a median of two (range 0–5) areas of focally increased radioactivity were seen at the proximal anastomosis (n = 21), in the body of the graft (n = 20) or at the distal anastomosis (n = 9). The activity was moderate in 27 cases and intense in 23 cases. Scintigraphic evidence of focal platelet aggregation in vein grafts was not correlated with preoperative antiplatelet therapy or vein graft diameter. Only 2 of the 20 intragraft platelet depositions occurred in areas where intraoperative vascular injury was suspected. In the composite graft and the PTFE grafts, diffuse activity was observed throughout the entire bypass. In conclusion, focal activity accumulations, suggesting localized endothelial injury, were observed in the majority of in situ vein bypasses, in particular at the sites of the anastomoses. Prosthetic bypasses were characterized by diffuse platelet aggregation.
Influence of body size and age on mitral ring motion
Authors: Wandt B. , Bojö L. , Wranne B.
Published: December 2004

Left ventricular systolic and diastolic function can be assessed by studying mitral ring motion. Reference values for a wide range of age and body size are lacking however. The motion of the mitral ring was studied with M‐mode echocardiography using the apical four‐ and two‐chamber views. Data are reported as the average of measurements of four points on the mitral ring. Data were analysed using the stepwise multiple regression technique, with age, gender, height, weight, body surface area and heart rate as independent variables. A total of 70 healthy subjects were studied. In children and adolescents under age 18, there was strong correlation between mitral ring motion and body size, age and heart rate. The ring motion was best described as mitral ring motion (mm) = 2·2+0·078 × height (cm) (SEE = 1·0 mm). In adults, mitral ring motion was correlated with age, height and heart rate but not with weight or body surface area. Ring motion could be described from the following equations: mitral ring motion (mm) = 12·7–0·060 × age (years)+0·031 × height (cm) (SEE = 1·2) or, if only age is taken into account, mitral ring motion = 18·4–0·065 × age (SEE = 1·2). In both children and adolescents up to age 18 and in adults, atrial contribution correlated significantly and positively to age and inversely to heart rate but not to height, weight, body surface area or gender. Atrial contribution was best described by the equation: atrial contribution = 0·15+0·0039 × age (SEE = 0·027). Thus, age and body size influences mitral ring motion and should be taken into account when interpreting patient data.
Mutational spectrum in Usher syndrome type II
Authors: Ouyang XM , Yam D , Hejtmancik JF , Jacobson SG , Li AR , Du LL , Angeli S , Kaiser M , Balkany T , Liu XZ
Published: December 2004

Usher syndrome type II is an autosomal recessive disorder characterized by moderate to severe hearing impairment and progressive visual loss due to retinitis pigmentosa (RP). We carried out a mutation screening of the USH2A gene in 88 probands with Usher syndrome type II to determine the frequency of USH2A mutations as a cause for USH2. Six mutations, including 2299delG, 921‐922insCAGC, R334W, N346H, R626X, and N357T were identified, with 2299delG mutation being the most frequent (16.5% of alleles), accounting for 77.5% of the pathologic alleles. Thirty‐five percent (31/88) of the probands had a USH2A mutation. Nine of them carried two pathogenic mutations: six cases were homozygotes and three were compound heterozygotes. Twenty‐two probands (25%) were found to carry only single USH2A mutations. One new missense mutation (N357T) occuring within the laminin N‐terminal (type VI) domain of usherin was identified. Eight polymorphisms were found, five of which are novel. Our data support the view that the 2299delG is the most common mutation in USH2A.
A Proteasome Inhibitor Prevents Vascular Hypertrophy In Deoxycorticosterone Acetate‐Salt Hypertensive Rats
Authors: Takaoka Masanori , Ohkita Mamoru , Itoh Makoto , Kobayashi Yutaka , Okamoto Hisako , Matsumura Yasuo
Published: December 2004

1. In the present study, we investigated the potential of the proteasome inhibitor N‐benzyloxycarbonyl‐Ile‐Glu(O‐t‐Bu)‐Ala‐leucinal (PSI) to prevent vascular hypertrophy induced by deoxycorticosterone acetate (DOCA) and salt in rats.

2. Vehicle (35% ethanol, 35% polyethylene glycol and 30% saline solution)‐treated DOCA‐salt rats developed marked hypertension at 4 weeks. Morphological studies on the rats given vehicle showed aortic hypertrophy, with a significant increase in wall thickness, wall area and wall‐to‐lumen ratio. A significant decrease in vascular wall hypertrophy was observed in PSI (3 mg/kg)‐treated DOCA‐salt rats. In addition, a marked increase in aortic endothelin (ET)‐1 content was evident in vehicle‐treated DOCA‐salt rats compared with findings in sham‐operated rats. A significant attenuation of this increase occurred in PSI‐treated DOCA‐salt rats.

3. These results indicate that PSI can prevent the vascular hypertrophy in DOCA‐salt hypertensive rats and the effect is accompanied by suppression of ET‐1 production in the aorta. We suggest that a proteasome‐dependent proteolytic system has an important role in the development of vascular hypertrophy in cases of DOCA‐salt‐induced hypertension, possibly through the enhancement of ET‐1 production in vascular tissues.