bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic analysis of the 2q33 region containing CD28–CTLA4–ICOS genes: association with non‐Hodgkin's lymphoma
Authors: Piras G. , Monne M. , Uras A. , Palmas A. , Murineddu M. , Arru L. , Bianchi A. , Calvisi A. , Curreli L. , Gaviano E. , Lai P. , Murgia A. , Latte G. C. , Noli A. , Gabbas A.
Published: June 2005

There is strong evidence that altered immunological function entails an increased risk of lymphoma, although the current knowledge of aetiological factors for lymphomas is limited. The CTLA4 gene encodes a receptor that provides a negative signal to the T‐cell once an immune response is initiated and completed. We analysed the 2q33 chromosomal region harbouring CD28, CTLA4 and ICOS genes, which are closely linked and have related functions in immune regulation, for association in 100 non‐Hodgkin's lymphoma (NHL) patients and in 128 healthy controls; both groups originated from Sardinia. There was a strong association of the CTLA4 49A and the 3′‐untranslated region (AT)82 alleles with NHL [odds ratio (OR) = 2, 95% confidence interval (CI) = 1·2–3·2, and OR = 1·6, 95% CI = 1·1–2·4 respectively]. CTLA4‐318C:49A:(AT)82 was the most represented haplotype in the studied population and was associated with NHL (P = 0·0029, OR = 1·76, 95% CI = 1·2–2·5). Strong linkage disequilibrium was detected between CD28, CTLA4 and ICOS and a ‘common’ haplotype was found very frequently among NHLs. However, no independent association between CD28, ICOS, D2S72 markers and NHL was observed. Our findings enable CTLA4 from adjacent functionally related genes as the true causative risk gene for NHL susceptibility at least in Sardinian patients.
Incidence of thrombotic complications in patients with haematological malignancies with central venous catheters: a prospective multicentre study
Authors: Cortelezzi Agostino , Moia Marco , Falanga Anna , Pogliani Enrico M. , Agnelli Giancarlo , Bonizzoni Erminio , Gussoni Gualberto , Barbui Tiziano , Mannucci Pier Mannuccio ,   the CATHEM Study Group
Published: June 2005

This prospective, observational and multicentre study assessed the incidence of, and risk factors for, symptomatic venous thrombotic complications after central venous catheter (CVC) positioning in patients with haematological malignancies. A total of 458 consecutive CVC insertions were registered in 416 patients (81·2% of whom had severe thrombocytopenia). Over the observation period (3 months or up to catheter removal), the incidence of events was: CVC‐related deep vein thrombosis (DVT), 1·5%; lower limb DVT, 0·4%; pulmonary embolism (PE), 1·3%; fatal PE, 0·6%; CVC‐related superficial thrombophlebitis, 3·9%; CVC‐occlusion/malfunction of thrombotic origin, 6·1%; major arterial events, 1·1%. Severe bleeding and CVC‐related infections were observed in 3·5% and 4·6% of cases respectively. A composite end point (any venous thromboembolism or superficial thrombophlebitis or CVC occlusion/malfunction) was defined in order to consider venous thrombotic events with a significant impact on clinical practice. With this criterion, the overall incidence was 12·0% (2·54 cases/1000 catheter days). No factor helped to predict venous thrombotic complications: only thrombocytopenia was associated with a weak trend for a reduced risk (odds ratio 0·52; 95% confidence interval 0·26–1·07). No severe bleeding was observed in those patients who received antithrombotic prophylaxis. This study shows that the impact on clinical practice of symptomatic CVC‐related thrombotic complications is not negligible in patients with haematological malignancies.
Outcomes of patients with acute leukaemia who relapsed after reduced‐intensity stem cell transplantation from HLA‐identical or one antigen‐mismatched related donors
Authors: Kobayashi Kazuhiko , Kami Masahiro , Murashige Naoko , Kusumi Eiji , Kishi Yukiko , Hamaki Tamae , Hori Akiko , Matsumura Tomoko , Yuji Koichiro , Masuo Shigeru , Mori Shinichiro , Miyakoshi Shigesaburo , Tanosaki Ryuji , Mitamura Tadayuki , Takaue Yoichi , Taniguchi Shuichi ,   the Tokyo SCT Consortium Institution
Published: June 2005

The characteristics of relapse following reduced‐intensity stem‐cell transplantation (RIST) remain to be clarified. We reviewed the medical records of 19 patients with acute leukaemia [acute myeloid leukaemia (AML), 16; acute lymphoblastic leukaemia (ALL), 3] who relapsed after RIST from related donors using purine‐analogue‐based regimens. Their median age was 55 years (range, 29–65 years). Median interval between RIST and relapse was 4·9 months (range, 1·8–24·9 months). Three chose not to receive interventions. The remaining 16 patients received withdrawal of immunosuppression (n = 3), chemotherapy (n = 2), donor lymphocyte infusion (n = 10) and second transplantation (n = 7), alone (n = 9) or in combination (n = 7). Four are alive with a median follow‐up of 27·6 months (range, 16·0–28·9 months); three in remission and one in relapse. The 2‐year overall survival after relapse was 28·9%. Causes of death in 15 patients included progressive disease (n = 7), graft‐versus‐host disease (n = 5) and infections (n = 3). Cumulative incidences of relapse‐related and non‐relapse‐related deaths at 2 years after relapse were 37% and 32% respectively. Two prognostic factors were identified on univariate analysis: age [P = 0·017; hazard ratio (HR), 1·16; 95% confidence interval (CI), 1·03–1·32], and ALL as underlying disease (P = 0·011; HR, 10·4; 95% CI, 1·73–62·4). Some AML patients who relapse after RIST achieve durable remission with allogeneic immunotherapy‐based interventions; however they carry a significant risk of non‐relapse mortality.
Genetic analysis in FXI deficiency: six novel mutations and the use of a polymerase chain reaction‐based test to define a whole gene deletion
Authors: Hill Marian , McLeod Fiona , Franks Hester , Gordon Ben , Dolan Gerry
Published: June 2005

The genetic basis of factor XI (FXI) deficiency was investigated in 30 patients from 13 different families of non‐Jewish origin. Twelve different mutations were detected (including six novel changes), seven missense mutations and three mutations leading to null alleles. Haplotype analysis suggested a large gene deletion in one family. We confirmed the presence of a recently reported Alu‐mediated FXI gene deletion. An unrelated patient with severe deficiency was shown to be compound heterozygous for A412V and this whole gene deletion. We suggest that this recurrent gene deletion should be included in the genetic analysis of FXI deficiency.
Cryoglobulin‐induced cytokine production via FcγRIIa: inverse effects of complement blockade on the production of TNF‐α and IL‐10. Implications for the growth of malignant B‐cell clones
Authors: Mathsson Linda , Tejde Andreas , Carlson Kristina , Höglund Martin , Nilsson Bo , Nilsson‐Ekdahl Kristina , Rönnelid Johan
Published: June 2005

Monoclonal antibodies produced by patients with lymphoproliferative diseases sometimes appear as cryoglobulins (CG), immunoglobulins (Ig) that reversibly agglutinate and form immune complexes (IC) when cooled below normal body temperature or through variation in pH and ionic strength. In accordance with our findings of IC‐induced cytokine production from peripheral blood mononuclear cells (PBMC) in systemic lupus erythematosus, we investigated whether CG can also induce cytokine production. One IgG and one IgM type I CG from two patients with multiple myeloma and Waldenstrom's macroglobulinaemia were individually purified and added to PBMC cultures. In separate experiments temperature and ionic strength were varied, or FcγRIIa, FcγRIII and complement activation were blocked; supernatant cytokine levels were then determined by enzyme‐linked immunosorbent assay. CG‐induced cytokine production from monocytes varied with precipitation induced by changes in temperature and ionic strength and was mediated via FcγRIIa‐ and complement‐dependent mechanisms. Complement blockade resulted in increased IgG CG‐induced interleukin (IL)‐10 production that was inversely correlated with decreased production of tumour necrosis factor‐α. CG‐induced IL‐10 might be a growth factor for malignant B‐lymphocytes in CG‐associated lymphoproliferative diseases with constant complement consumption. Knowledge of mechanisms underlying CG‐induced cytokine production can be useful for designing treatments for type I CG‐associated pathology in lymphoproliferative diseases.
Bortezomib therapy alone and in combination with dexamethasone for previously untreated symptomatic multiple myeloma
Authors: Jagannath Sundar , Durie Brian G. M. , Wolf Jeffrey , Camacho Elber , Irwin David , Lutzky Jose , McKinley Marti , Gabayan Eli , Mazumder Amitabha , Schenkein David , Crowley John
Published: June 2005

Bortezomib, as a single agent and in combination with dexamethasone, was examined as first‐line treatment in 32 consecutive patients with untreated symptomatic multiple myeloma. Patients received bortezomib 1·3 mg/m2 for a maximum of six 3‐week cycles; oral dexamethasone 40 mg was added if a less than partial response (PR) was achieved after two cycles or a less than complete response (CR) was achieved after four cycles. The response rate (CR + PR) was 88%, with undetectable paraprotein (CR) in 6%, and detectable by immunofixation only in 19% [near (n)CR]. All 32 patients completed the first two cycles of bortezomib alone, of whom 3% achieved CR, 9% nCR, and 28% PR. Ten patients received single‐agent bortezomib on study, and dexamethasone was added in 22, leading to 15 improved responses. The most common adverse events ≥grade 2 included sensory neuropathy (31%), constipation (28%), myalgia (28%) and fatigue (25%). Sensory neuropathy grade 2 or 3 was reversible within a median of 3 months in five of 10 patients. Bortezomib treatment did not affect stem cell mobilization in eight or transplantation in six patients. Bortezomib alone or in combination with dexamethasone is an effective induction therapy with a high CR and nCR rate and manageable toxicities in previously untreated patients with myeloma.
LISA: a web‐based decision‐support system for trial management of childhood acute lymphoblastic leukaemia
Authors: Bury Jonathan , Hurt Chris , Roy Anindita , Cheesman Louise , Bradburn Mike , Cross Simon , Fox John , Saha Vaskar
Published: June 2005

Continuation chemotherapy is a key component of the treatment of childhood acute lymphoblastic leukaemia. During this treatment phase, weekly dose adjustments are carried out based on current and historical full blood counts (FBCs). The dose decision pathway is complex and suboptimal therapy may result if information on FBC results is not readily available and/or the prescriber is inexperienced. A web‐based decision‐support system (Leukaemia Intervention Scheduling and Advice, ‘LISA’) was designed to facilitate access to FBC information across geographical locations and to assist with dosage adjustments. A balanced‐block crossover analysis was performed to evaluate the system. Thirty‐six clinicians with varying degrees of experience were each asked to decide on appropriate oral chemotherapy dosages for eight simulated cases: four using LISA and four without. LISA significantly reduced the number of erroneous prescriptions (zero of 144 with LISA vs. 54 of 144 without; P < 0·0001) without affecting the number of times subjects deliberately overrode the protocol (seven of 144 times using LISA and six of 144 without). Using LISA reduced the time taken by novices to reach a decision for each case but increased the time taken by experts. Thirty‐five of 36 subjects said they would be likely to use the system if it were available. A system like LISA is likely to be acceptable to clinicians, and has the potential to increase protocol compliance and decrease prescribing errors while allowing clinicians to override the protocol in specific cases where sound reasons exist for doing so.
Prevalence of adrenal androgen excess in patients with the polycystic ovary syndrome (PCOS)
Authors: Kumar Ashim , Woods Keslie S. , Bartolucci Alfred A. , Azziz Ricardo
Published: June 2005

Objective  To determine the prevalence of adrenal androgen (AA) excess in the polycystic ovary syndrome (PCOS) using age‐ and race‐specific normative values.

Design  Cross‐sectional observational study.

Patients  One hundred and eight‐two (88 Black and 94 White) age‐matched healthy eumenorrhoeic nonhirsute women (controls) and 213 (27 Black and 186 White) women with PCOS were recruited.

Measurements  Total testosterone (T), free T, androstenedione (A4), dehydroepiandrosterone sulfate (DHEAS) and SHBG, as well as fasting insulin and glucose, were measured in plasma.

Results  The mean total T, free T, A4, DHEAS and body mass index (BMI) were higher in women with PCOS than in control women. DHEAS levels were significantly lower in Black controls than White controls, whereas fasting insulin and BMI were higher in Black controls. In control and Black PCOS women, DHEAS levels did not correlate with BMI, waist‐to‐hip ratio (WHR) or fasting insulin. Among White women with PCOS, DHEAS levels correlated negatively with BMI and fasting insulin. DHEAS levels decreased similarly with age in control and PCOS women of either race. For each race and age group the upper 95% normative values for log DHEAS was calculated, and the number of PCOS subjects with log DHEAS values above this level were assessed. The prevalence of supranormal DHEAS levels was 33·3% and 19·9%, respectively, among Black and White women with PCOS.

Conclusions  The prevalence of DHEAS excess is approximately 20% among White and 30% among Black PCOS patients, when using age‐ and race‐adjusted normative values. This study also indicates that the age‐associated decline in DHEAS levels is observable and similar in both control and PCOS women, regardless of race. While BMI and fasting insulin had little impact on circulating DHEAS levels in healthy women, among White PCOS patients these parameters were negatively associated with circulating DHEAS levels.
Growth hormone secretion in adult patients with thalassaemia
Authors: La Rosa Clementina , De Sanctis Vincenzo , Mangiagli Antonino , Mancuso Michele , Guardabasso Vincenzo , Galati Maria Concetta , Caruso‐Nicoletti Manuela
Published: June 2005

Background and Objectives  Growth retardation and short stature are frequent clinical features of patients with beta‐thalassaemia major. Dysfunction of the GH–IGF‐1 axis has been described in many thalassaemic children and adolescents with short stature and reduced growth velocity. Several studies have demonstrated that recombinant GH treatment improves growth velocity in these patients, although response to the treatment is variable and not predictable. A reassessment of the GH–IGF‐1 axis must be performed in young adults with childhood‐onset GH deficiency (GHD), after attainment of final height, to select those who are candidates for replacement therapy as adults. To our knowledge there are no data available on retesting the GH–IGF‐1 axis in adult thalassaemic patients with childhood‐onset GHD. The aim of our study was to investigate GH secretion in adult thalassaemic patients with childhood‐onset GHD.

Design  We reassessed GH secretion in a group of adult thalassaemic patients in whom partial GHD had been diagnosed during childhood.

Patients and Methods  We performed an arginine plus GH‐releasing hormone (GHRH) stimulation test in 16 thalassaemic patients (10 males, six females) with a mean age of 24·8 ± 3·6 years. The cut‐off level for GH response was set at 9 µg/l, according to the literature. Ferritin, IGF‐1, liver enzymes and lipid levels were also determined.

Results  We found persisting GHD in three patients, one patient had borderline values (GH peak = 10·4 µg/l), whereas the others had a normal response. These results are in accordance with the data on GH retesting in adult patients with idiopathic partial childhood‐onset GHD.

Conclusion  We conclude that GH status should be retested in adult thalassaemic patients with childhood‐onset GHD. If the diagnosis of adult GHD is established, GH treatment may be considered as it could contribute to improve heart function and bone mineral density, which are frequently impaired in adult thalassaemic patients.
Decreased type 1 iodothyronine deiodinase expression might be an early and discrete event in thyroid cell dedifferentation towards papillary carcinoma
Authors: De Souza Meyer Erika L. , Dora José Miguel , Wagner Márcia S. , Maia Ana Luiza
Published: June 2005

Objective  Type I iodothyronine deiodinase (D1) catalyses the 5′ monodeiodination of T4 and is highly expressed in normal human thyroid gland. We have investigated D1 expression in a series of benign and malignant differentiated thyroid neoplasias.

Design  Surgically isolated thyroid tumour fragments were used. D1 expression was determined by reverse transcription polymerase chain reaction (RT‐PCR) and enzymatic assay.

Patients  Tumours and adjacent normal tissues were obtained from 28 unselected patients (papillary carcinoma, n = 14; follicular adenoma, n = 7; follicular carcinoma, n = 6; anaplastic carcinoma, n = 1).

Measurements  D1 mRNA levels were determined using specific primers for the human D1 gene and enzymatic assays were performed using T4 as substrate.

Results  In papillary thyroid carcinoma (PTC), D1 mRNA and activity levels were decreased compared with the surrounding tissue (0·25 ± 0·24 vs. 1·09 ± 0·54 arbitrary units (AU), P < 0·001 and 0·08 ± 0·07 vs. 0·24 ± 0·15 pmol T4/min/mg protein, P = 0·045, respectively). Decreased D1 expression was consistent and was observed in all histological subtypes and clinical stages analysed, including microcarcinomas. By contrast, significantly higher D1 mRNA levels and enzyme activity were present in follicular adenoma (1·9 ± 1·5 vs. 0·83 ± 0·58 AU, P = 0·028 and 2·67 ± 1·42 vs. 0·22 ± 0·06 pmol T4/min/mg protein, P = 0·044, respectively) and in follicular thyroid carcinoma (FTC) than in surrounding normal tissue (1·2 ± 0·46 vs. 0·67 ± 0·18 AU, P = 0·038 and 1·20 ± 0·58 vs. 0·20 ± 0·10 pmol T4/min/mg protein, P < 0·001, respectively). Type II iodothyronine deiodinase (D2) activity was also significantly higher in metastatic FTC samples than in normal thyroid tissues (5·20 ± 0·81 vs. 0·30 ± 0·27 fmol T4/min/mg protein, P < 0·001).

Conclusions  These findings suggest that thyroid cell dedifferentiation promotes changes in D1 gene expression by pretranscriptional mechanisms and indicate that decreased D1 expression might be an early and discrete event in thyroid cell dedifferentiation towards papillary thyroid carcinoma.
The nadir growth hormone after an octreotide test dose predicts the long‐term efficacy of somatostatin analogue therapy in acromegaly
Authors: Gilbert J. A. , Miell J. P. , Chambers S. M. , McGregor A. M. , Aylwin S. J. B.
Published: June 2005

Objective  In the treatment of acromegaly, a ‘test dose’ of octreotide is recommended prior to the use of depot somatostatin analogue (SSA) therapy. However, there remains no consensus regarding the criteria that predict a response to treatment. The ability to select patients who may benefit most from medical therapy is potentially of great value in clinical practice. The aim of the study was to determine the predictive value of both the nadir GH and the mean GH following an octreotide test dose in identifying patients who subsequently achieved disease remission with depot SSA therapy. Remission was defined as a mean GH < 5 mU/l (< 2 µg/l).

Design  Retrospective case–control study.

Patients  A group of 41 patients with acromegaly underwent an octreotide test dose where GH was measured hourly for a total of 6 h following an injection of octreotide 50 µg subcutaneously. Nadir GH and mean GH following the octreotide test dose were determined. Thirty‐three patients were subsequently treated with depot SSA therapy and mean GH and IGF‐I levels were determined at follow‐up.

Results  The nadir GH demonstrated superior predictive power to that of mean GH across a range of GH cut‐off values. A nadir GH < 5 mU/l demonstrated 80% sensitivity and 83% specificity in predicting remission with depot SSA therapy. A nadir GH < 10 mU/l demonstrated 100% sensitivity and 56% specificity.

Conclusions  The nadir GH following an octreotide test dose is a useful predictive marker of achieving disease remission with depot SSA therapy used as either a primary or an adjuvant agent.
Lymphoid tyrosine phosphatase (PTPN22/LYP) variant and Graves’ disease in a Polish population: association and gene dose‐dependent correlation with age of onset
Authors: Skórka Agata , Bednarczuk Tomasz , Bar‐Andziak Ewa , Nauman Janusz , Ploski Rafal
Published: June 2005

Objective  Susceptibility to Graves’ disease (GD) is to a significant extent determined by genetic factors of which the best known are those associated with the HLA and the CTLA4 locus. Recently, two studies on British Caucasians reported that a single nucleotide polymorphism, 1858 C>T in PTPN22, encoding Arg620Trp in the lymphoid protein tyrosine phosphatase (LYP), which is a negative regulator of T‐cell activation, increases the risk of GD. The purpose of our study was to investigate whether the PTPN22‘T’ allele is associated with GD and/or its subsets, defined by clinical or genetic parameters, in a Polish population.

Subjects and design  A cohort of 290 patients and 310 controls was genotyped using a PCR‐RFLP method. The distribution of PTPN22 alleles and genotypes among patients and controls was compared, and correlation was sought between PTPN22‘T’ and sex, tobacco smoking status, family history of GD, age of disease onset, presence (and severity) of ophthalmopathy, and presence of the CTLA4 A49G or DRB1*03 alleles.

Results  Association between GD and the PTPN22‘T’ allele was confirmed (OR 1·7, P < 0·0008). Furthermore, a significant correlation between the PTPN22 genotype and the age of GD onset was demonstrated (r = −0.18, P = 0·0019). The PTPN22‘TT’ and ‘CC’ genotypes defined groups characterized by more than twofold difference in median age of disease onset (20·8 years vs. 42 years, P < 0·003) whereas the ‘CT’ genotype was associated with an intermediate value (35 years). There were no statistically significant correlations with other analysed clinical or genetic parameters.

Conclusions  We replicated the association between Graves’ disease and PTPN22‘T’ reported in British Caucasians. We also found a gene dose‐dependent effect of PTPN22‘T’ on the age of onset of Graves’ disease.
Circulating cortisone levels are associated with biochemical markers of bone formation and lumbar spine BMD: the Hertfordshire Cohort Study
Authors: Cooper Mark S. , Syddall Holly E. , Fall Caroline H. D. , Wood Peter J. , Stewart Paul M. , Cooper Cyrus , Dennison Elaine M.
Published: June 2005

Objective  Cortisone is an endogenous corticosteroid that has negligible intrinsic glucocorticoid activity but can be converted to the active corticosteroid cortisol by the enzyme 11β‐hydroxysteroid dehydrogenase type 1 (11β‐HSD1). 11β‐HSD1 is expressed in osteoblasts and may play a role in determining susceptibility to glucocorticoid‐induced osteoporosis. In intact osteoblasts enzyme activity, and thus cortisol generation, is dependent on substrate concentration with an almost linear increase in activity across the physiological range. We have therefore attempted to measure the impact of 11β‐HSD1 activity on bone in vivo by examining the association of circulating cortisone with bone markers, bone mineral density (BMD) and bone loss in a cohort of women and men.

Design and subjects  Baseline cross‐sectional association study involving 135 women and 171 men aged 61–73 years from the Hertfordshire Cohort Study and a 4 year follow‐up study examining changes in BMD.

Measurements  Serum cortisone, cortisol and osteocalcin, and urinary type I collagen cross‐linked N‐telopeptide (NTX) were measured at baseline. BMD at spine and hip was measured at baseline and 4 years later.

Results  In men serum cortisone levels were negatively correlated with serum osteocalcin (r = −0·20, P = 0·01); a similar relationship was seen in women (r = −0·16, P = 0·06). No correlation was seen between serum cortisone and urinary NTX (r = 0·03, P = 0·74 for women; r = −0·03, P = 0·72 for men). A negative correlation was observed between serum cortisone and spine BMD in women (r = −0·18, P = 0·04); a similar relationship was also seen in men (r =−0·14, P = 0·07). However, cortisone did not correlate with BMD at the femoral neck or total hip or changes in BMD at any site over time. In analyses adjusted for adiposity, osteoarthritis grade and a range of life‐style variables, these relationships did not change substantially. All these relationships were independent of cortisol concentrations.

Conclusions  The most plausible explanation for the association of circulating cortisone levels with osteocalcin is the presence of 11β‐HSD1 activity within osteoblasts. The measurement of serum cortisone may independently give insights into the action of glucocorticoids on bone.
The effects of 12 months of growth hormone replacement therapy on cardiac autonomic tone in adults with growth hormone deficiency
Authors: Tanriverdi F. , Eryol N. K. , Atmaca H. , Unluhizarci K. , Ozdogru I. , Sarikaya I. , Bayram F. , Kelestimur F.
Published: June 2005

Objectives  Growth hormone deficiency (GHD) in adults is associated with a cluster of cardiovascular risk factors. Some abnormalities of cardiac structure and function have been reported in adult patients with GHD, but there are few data related to cardiac autonomic tone. Non‐invasive assessment of cardiac autonomic status can be achieved by heart rate variability (HRV), which can be measured by using time‐domain or frequency‐domain variables. To our knowledge, short‐term (6 months) effects of GH replacement therapy (GHRT) on HRV in a limited number of patients have been evaluated prospectively in only two previous studies. The present study was therefore designed to investigate the effects of GHD and 12 months of GHRT on cardiac autonomic tone in a larger number of adult patients with severe GHD.

Patients and methods  HRV measurement, by using time‐domain variables, was performed in 22 patients with GHD (eight men, 14 women; mean age 45·4 ± 2·4 years) and 22 healthy controls (nine men, 13 women; mean age 40·8 ± 1·8 years) at baseline. The time‐domain variables (sympathetically influenced parameters SDNN and SDANN and parasympathetically influenced parameters RMSSD and PNN50) were derived from 24‐h electrocardiogram (ECG) recordings. In the patient group, cardiac autonomic tone was re‐evaluated after 6 and 12 months of GHRT.

Results  Mean baseline values of SDNN and SDANN were significantly higher (higher values mean lower sympathetic activity) in GHD patients than in healthy controls (P < 0·05), but mean baseline values of RMSSD and PNN50 did not differ significantly in healthy controls and patients. After 6 and 12 months of GHRT, mean SDNN and SDANN were decreased significantly when compared with the baseline values before GHRT (P < 0·05). However, mean RMSSD and PNN50 did not differ significantly from baseline. When SDNN and SDANN measurements were evaluated individually for each patient, after 12 months of GHRT both of the sympathetically influenced parameters decreased in 90% of the patients.

Conclusions  These data indicate that sympathetic tone is decreased in adult patients with severe GHD. Additionally, an increment in sympathetic activity and normalization of sympathovagal balance have been demonstrated after 6 and 12 months of GHRT. This result suggests that, at least at the doses used in this study, GHRT improves sympathetic tone, without an obvious arrhythmogenic effect.
A population study of the association between thyroid autoantibodies in serum and abnormalities in thyroid function and structure
Authors: Bülow Pedersen Inge , Laurberg Peter , Knudsen Nils , Jørgensen Torben , Perrild Hans , Ovesen Lars , Rasmussen Lone Banke
Published: June 2005

Objective  Patients with autoimmune overt hypothyroidism may present with goitrous Hashimoto's disease or autoimmune atrophic thyroiditis. Little is known about the prevalence of subclinical autoimmune hypothyroidism. The aims of this study were to evaluate the association between thyroid autoantibodies in serum and abnormalities in thyroid function and structure, and to study the thyroid volume in subjects with subclinical autoimmune hypothyroidism.

Design  A population study including 4649 randomly selected subjects.

Measurements  Blood tests were used to analyse for thyroid peroxidase autoantibodies (TPO‐Ab), thyroglobulin autoantibodies (Tg‐Ab), TSH, fT3 and fT4.

Results  Thyroid volume was categorized as small (< 6·6 ml) in 4·7%, normal (6·6–14·9 ml) in 60·4% and large (> 14·9 ml) in 34·9% of participants. Thyroid nodules were found in 29·7%. Serum TSH was low (< 0·4 mIU/l) in 4·7%, normal (0·4–3·6) in 91·0% and high (> 3·6) in 4·3%. The prevalence rate of subclinical goitrous Hashimoto's disease was 0·62% and of subclinical autoimmune atrophic thyroiditis 0·24%. There was a strong association between large volume and autoantibodies, but only in subjects with elevated TSH (P < 0·001). An association between thyroid nodules and TPO‐Ab in univariate analyses (P < 0·001) was due to confounding by sex and age (multivariate model, P = 0·23).

Conclusion  We identified a subgroup of the population with subclinical goitrous Hashimoto's disease and a smaller subgroup with subclinical autoimmune atrophic thyroiditis. This relationship between small and large thyroid volume in subclinical disease is opposite to that in overt disease, which may suggest that the period between development of a small volume with circulating autoantibodies and overt hypothyroidism is relatively short.
The metabolic syndrome in middle‐aged men according to different definitions and related changes in carotid artery intima‐media thickness (IMT) during 3 years of follow‐up
Authors: WALLENFELDT K. , HULTHE J. , FAGERBERG B.
Published: June 2005

Objectives.  To examine the occurrence over time of the metabolic syndrome (MetS) according to different definitions and the relation to change during follow‐up in carotid artery intima‐media thickness (IMT), measured by ultrasound.

Design.  A cohort of 316, originally 58‐year‐old men, initially free of diabetes and cardiovascular disease, was followed for 3.2 ± 0.2 years. IMT was measured bilaterally by high‐resolution B‐mode ultrasound at baseline and follow‐up. The MetS was classified according to slightly modified World Health Organization (WHO) and National Cholesterol Education Program (NCEP) criteria.

Results.  In 88% WHO and NCEP definitions resulted in identical classifications. IMT was larger both at baseline and after 3 years in men fulfilling the criteria for the MetS, according to either of the definitions, compared to those without factors in the syndrome. Men who fulfilled the WHO criteria for the MetS, at the initial and final examination showed a statistically significant increase in carotid artery IMT during the study [76 (95% CI: 14–130) μm, n = 37]. Men fulfilling the WHO criteria for the MetS at baseline tended to have a larger annual increase in IMT than those not fulfilling the criteria or having no risk factors in the syndrome.

Conclusions.  More than 10% of the men had the MetS both at baseline and after 3 years, and this was associated with an increase in IMT using the WHO definition. Several of the components included in the MetS deteriorated during follow‐up, i.e. body mass index (BMI), waist‐to‐hip ratio (WHR), waist circumference, blood glucose and blood pressure.
Glucocorticoid Enhances the Neurotoxic Actions of Quinolinic Acid in the Striatum in a Cell‐Specific Manner
Authors: Ngai L.‐Y. , Herbert J.
Published: June 2005

This study demonstrates that corticosterone can exacerbate the damaging effects of infused quinolinic acid (QA) on the dorsal striatum. Adult adrenalectomised male rats were pretreated subcutaneously with graded doses of corticosterone (0, 0.5, 2, 5, 20 and 40 mg/kg/day) for 2 days and then received a unilateral infusion of QA (45 nmol) (under Isoflurane/N2O anaesthesia) into the dorsal striatum. A control infusion (vehicle) was made into the striatum on the other side. Corticosterone treatment was continued and they were killed 7 days later. Plasma corticosterone was measured by radioimmunoassay, and thymus weights were used as an integrated measure of glucocorticoid activity. Lesion volumes were measured on neuronal nuclei stained sections, dopamine and cyclic AMP‐regulated phosphoprotein 32 (DARRP‐32) was used to assess medium spiny neurone survival, NADPH‐diaphorase histochemistry to assess medium aspiny neurones and, finally, choline acetyltransferase to assess large aspiny neurones. Adrenalectomised rats showed smaller lesions than control (sham‐operated) rats, suggesting significant protection. Increasing doses of corticosterone resulted in larger lesions up to an apparent ceiling effect at higher doses; there was no evidence of a U‐shaped dose–response. There was a differential effect of both QA and corticosterone on the cell populations of the striatum. Medium spiny neurones were most vulnerable to the effects of QA and to the exacerbating actions of corticosterone. Medium aspiny neurones were equally vulnerable to QA but corticosterone had no additional effect. Large aspiny neurones were relatively less sensitive to QA and there was no additional action of administered corticosterone. These results show that corticosterone has a selective neuroendangering action within the striatum, but there is no evidence for a protective action of glucocorticoids at lower doses.
Characterization of Chlamydia MurC‐Ddl, a fusion protein exhibiting D‐alanyl‐D‐alanine ligase activity involved in peptidoglycan synthesis and D‐cycloserine sensitivity
Authors: McCoy Andrea J. , Maurelli Anthony T.
Published: June 2005

Recent characterization of chlamydial genes encoding functional peptidoglycan (PG)‐synthesis proteins suggests that the Chlamydiaceae possess the ability to synthesize PG yet biochemical evidence for the synthesis of PG has yet to be demonstrated. The presence of D‐amino acids in PG is a hallmark of bacteria. Chlamydiaceae do not appear to encode amino acid racemases however, a D‐alanyl‐D‐alanine (D‐Ala‐D‐Ala) ligase homologue (Ddl) is encoded in the genome. Thus, we undertook a genetics‐based approach to demonstrate and characterize the D‐Ala‐D‐Ala ligase activity of chlamydial Ddl, a protein encoded as a fusion with MurC. The full‐length murC‐ddl fusion gene from Chlamydia trachomatis serovar L2 was cloned and placed under the control of the arabinose‐inducible ara promoter and transformed into a D‐Ala‐D‐Ala ligase auxotroph of Escherichia coli possessing deletions of both the ddlA and ddlB genes. Viability of the E. coliΔddlAΔddlB mutant in the absence of exogenous D‐Ala‐D‐Ala dipeptide became dependent on the expression of the chlamydial murC‐ddl thus demonstrating functional ligase activity. Domain mapping of the full‐length fusion protein and site‐directed mutagenesis of the MurC domain revealed that the structure of the full fusion protein but not MurC enzymatic activity was required for ligase activity in vivo. Recombinant MurC‐Ddl exhibited substrate specificity for D‐Ala. Chlamydia growth is inhibited by D‐cycloserine (DCS) and in vitro analysis provided evidence for the chlamydial MurC‐Ddl as the target for DCS sensitivity. In vivo sensitivity to DCS could be reversed by addition of exogenous D‐Ala and D‐Ala‐D‐Ala. Together, these findings further support our hypothesis that PG is synthesized by members of the Chlamydiaceae family and suggest that D‐amino acids, specifically D‐Ala, are present in chlamydial PG.
Genetic analysis of the Legionella pneumophila DotB ATPase reveals a role in type IV secretion system protein export
Authors: Sexton Jessica A. , Yeo Hye‐Jeong , Vogel Joseph P.
Published: June 2005

The pulmonary pathogen Legionella pneumophila uses the Dot/Icm type IV secretion system (T4SS) to replicate inside host cells. This apparatus translocates proteins into macrophages to alter their endocytic pathway and enable bacterial growth. Although the secretion ATPase DotB is critical for T4SS function, its specific role in type IV secretion remains undefined. Due to similarity to the VirB11 and PilT ATPases, DotB has been proposed to play a role in assembly of the T4SS, retraction of pili and/or export of substrates. With the goal of understanding the protein's function(s), we isolated and characterized 30 dotB alleles using a variety of phenotypic and biochemical assays. Twenty‐four of these alleles possess several dot/icm mutant phenotypes, including a complete lack of intracellular replication, plasmid mobilization and contact‐dependent cytotoxicity. These 24 non‐functional alleles fall into three classes: those with a known biochemical defect, those with a predicted enzymatic defect and those with an unknown defect. Six other alleles display partial activity in dot/icm phenotypic assays, thus constituting a fourth class. Two mutants in this class are unable to export a subset of T4SS substrates, providing the first evidence for a DotB function in substrate export and suggesting a possible role in substrate selection.
The Aspergillus nidulans sldIRAD50 gene interacts with bimEAPC1, a homologue of an anaphase‐promoting complex subunit
Authors: Malavazi Iran , Lima Joel Fernandes , Fagundes Márcia Regina von Zeska Kress , Efimov Vladimir P. , Goldman Maria Helena de Souza , Goldman Gustavo Henrique
Published: June 2005

The Mre11–Rad50–Nbs1 protein complex has emerged as a central component in the human cellular DNA damage response, and recent observations suggest that these proteins are at least partially responsible for the linking of DNA damage detection to DNA repair and cell cycle checkpoint functions. We have identified Aspergillus nidulans sldI1444D mutant in a screen for dynein synthetic lethals. The sldIRAD50 gene was cloned by complementation of the sporulation deficiency phenotype of this mutant. A transversion G→C at the position 2509 (Ala‐692‐Pro amino acid change) in the sldI1444D mutant causes sensitivity to several DNA‐damaging agents. The mutation sldI1 occurs at the CXXC hinge domain of Rad50. We have deleted part of the coiled‐coil and few amino acids of the Rad50–Mre11 interaction region and assessed several phenotypic traits in this deletion strain. Besides sensitivity to a number of DNA‐damaging agents, this deletion strain is also impaired in the DNA replication checkpoint response, and in ascospore viability. There is no delay of the S‐phase when germlings of both sldI RAD50 and mreAMRE11 inactivation strains were exposed to the DNA damage caused by bleomycin. Transformation experiments and Southern blot analysis indicate homologous recombination is dependent on scaANBS1 function in the Mre11 complex. There are epistatic and synergistic interactions between sldI RAD50 and bimEAPC1 at S‐phase checkpoints and response to hydroxyurea and UV light. Our results suggest a possible novel feature of the Mre11 complex in A. nidulans, i.e. a relationship with bimE APC1.
Identification of the membrane penetrating domain of Vibrio cholerae cytolysin as a β‐barrel structure
Authors: Valeva Angela , Walev Iwan , Boukhallouk Fatima , Wassenaar Trudy M. , Heinz Natalia , Hedderich Julia , Lautwein Sina , Möcking Moritz , Weis Silvia , Zitzer Alexander , Bhakdi Sucharit
Published: June 2005

Vibrio cholerae cytolysin (VCC) is an oligomerizing pore‐forming toxin that is related to cytolysins of many other Gram‐negative organisms. VCC contains six cysteine residues, of which two were found to be present in free sulphydryl form. The positions of two intramolecular disulphide bonds were mapped, and one was shown to be essential for correct folding of protoxin. Mutations were created in which the two free cysteines were deleted, so that single cysteine substitution mutants could be generated for site‐specific labelling. Employment of polarity‐sensitive fluorophores identified amino acid side‐chains that formed part of the pore‐forming domain of VCC. The sequence commenced at residue 311, and was deduced to form a β‐barrel in the assembled oligomer with the subsequent odd‐numbered residues facing the lipid bilayer and even‐numbered residues facing the lumen. Pro328/Lys329 were tentatively identified as the position at which the sequence turns back into the membrane and where the antiparallel β‐strand commences. This was deduced from fluorimetric analyses combined with experiments in which the pore was reversibly occluded by derivatization of sulphydryl groups with a bulky moiety. Our data support computer‐based predictions that the membrane‐permeabilizing amino acid sequence of VCC is homologous to the β‐barrel‐forming sequence of staphylococcal cytolysins and identify the β‐barrel as a membrane‐perforating structure that is highly conserved in evolution.
Memories of Biology, Mathematics and Ivan
Authors: Henry C.
Published: June 2005

The plaque assay years at Pittsburgh that included efforts to optimize the detection for single immunoglobulin M (IgM) and IgG antibody‐producing cells are described. Sensitivity of the assay and precision of the plaque count was established, and a series of response curves were obtained that enabled to calculate the frequency of precursors and to identify the target of the antigen. If one ignored a number of anomalies of the curves and made the simple assumption that a single hit by antigen on a pre‐committed precursor yielded a clone of antibody‐forming cells, estimates of some 700 target cells (per mouse spleen) were obtained, figures not too far from the number that Ivan would later find.

Years at the Paul Ehrlich Institute and visits to the Basel Institute for Immunology are described.
Pittsburgh 1962/63 Revisited: Too Many Antibodies, Too Few Ribosomes?
Authors: Staehelin T.
Published: June 2005

Jerne's success in developing the plaque‐forming cell methodology for determining the number of specific antibody‐producing lymphocytes during the immune response has prompted scientists to calculate the number of antibody molecules produced by a single cell. In this article, calculations of the rate of antibody production are made, and a special attention is called to the number of ribosomes in a cell, the rate of protein chain elongation and various other parameters. The generally accepted notion of 2000 IgM molecules or some 15,000 IgG molecules per second (per cell) remains an acceptable estimate. The article describes the work of Ivan Lefkovits and the interaction among research groups leading to quantitative evaluations of the components of the immune system.
From Poisson Distribution to Interleukins
Authors: Aarden L.
Published: June 2005

In the mid‐1970s, Ivan Lefkovits and co‐workers introduced limiting dilution analysis of T and B cells as a method to quantify antigen‐specific T and B cells. In this study, I describe how my co‐operation with Ivan on the subject led not only to heated discussions on the validity of the approach but also to the identification of interleukin‐2 as T‐cell‐derived factor, capable of replacing T‐cell help required for the activation of B cells.
Importance of Counting in Biology
Authors: Pink J. R. L.
Published: June 2005

In this study, analysis of biological problems through structural and quantitative approaches is described. Classical genetics on major histocompatibility complex in avian species has been evaluated. Experiments showing that antibodies from various individuals were generally distinct have led to further analysis of immunoglobulin light chain diversity by means of O'Farrell's 2D gel technique. The transition of the research towards vaccine development has been described.

   ‘It's not the voting that's democracy, it's the counting.'

  – Tom Stoppard (Czech‐born dramatist), Jumpers (1972) act 1.
Dissection of Systems, Cell Populations and Molecules
Authors: Benner R. , Khan N. A.
Published: June 2005

This paper summarizes studies on antibody formation in the bone marrow and the suppressive effects of intravenous immunization with allogeneic blood cells on T‐cell function in mice. The latter studies were extended by employing the limiting dilution culture system developed in Ivan Lefkovits' laboratory and implemented in collaboration with Lucien Aarden. Thereby, the functional data were complemented with frequencies of alloantigen‐activated helper (Th) and suppressor T cells after intravenous alloimmunization. These results led the Rotterdam group to studies on the prevention of rejection of the foetal ‘allograft’. Th cells are central in foetal allograft rejection and pregnancy success. Characteristic for human pregnancy is the production of the glycoprotein chorionic gonadotropin (hCG) hormone. The in vivo liberated peptide fragments originating from nicking of the sequence MTRVLQGVLPALPQ in the β‐chain of hCG were considered for their immunoregulating capacity related to pregnancy success. These peptides – prepared synthetically – (MTR, MTRV, LQG, LQGV, VLPALP and others) indeed showed a remarkable spectrum of biological effects (e.g. modulation of angiogenesis, inhibition of septic shock syndrome, prevention of diabetes and reduction of ischaemia‐reperfusion damage). The paper interprets and generalizes these findings and projects them into various research directions, especially towards the proteomics framework studies built up in Ivan Lefkovits' laboratory in the nineties. During the time period, when Ivan spent a mini‐sabbatical in Rotterdam (months after closing down the BII) more detailed discussions were intiated. This paper is meant to keep the discussions between the involved research groups going on.
Mouse Hepatitis Virus 3 Binding to Macrophages Correlates with Resistance to Experimental Infection
Authors: Pereira C. A. , Moreira C. , Tsuhako M. H. , De Franco M. T.
Published: June 2005

Mouse hepatitis virus 3 (MHV3) infection of A/J and BALB/c mice has been used as a model of resistance/susceptibility. A/J mice recover from a mild disease after 4–6 days of infection and the BALB/c mice develop an acute hepatitis and die after 3–4 days of infection. In view of studying the MHV3 binding to cells or cell extracts, we performed an enzyme‐linked immunosorbent assay‐like virus‐binding assay, preparing microplates with L929 cells, A/J or BALB/c mouse macrophages and also with proteins extracted from these cells. Higher MHV3 bindings were observed to proteins of BALB/c macrophages than to the A/J ones. The interferon‐γ (IFN‐γ) activation led to a reduction of MHV3 binding only to proteins of resistant A/J mouse macrophages. Our experiments contribute to the hypothesis that IFN‐γ activation of macrophages plays an important role against MHV3 infection by downregulating the expression of viral receptors.
Comparison of select innate immune mechanisms of fish and mammals
Authors: Plouffe Debbie A. , Hanington Patrick C. , Walsh John G. , Wilson Elaine C. , Belosevic Miodrag
Published: June 2005

Abstract:  The study of innate immunity has become increasingly popular since the discovery of homologs of many of the innate immune system components and pathways in lower organisms including invertebrates. As fish occupy a key position in the evolution of the innate and adaptive immune responses, there has been a great deal of interest regarding similarities and differences between their defense mechanisms and those of higher vertebrates. This review focuses on describing select mechanisms of the innate immune responses of fish and the implications for evolution of immunity in higher vertebrates.