bluuebunny/crossref_metadata_2025 huggingface dataset:
Subcutaneous adipose tissue expression of tumour necrosis factor‐α is not associated with whole body insulin resistance in obese nondiabetic or in type‐2 diabetic subjects
Authors: Koistinen  , Bastard  , Dusserre  , Ebeling  , Zegari  , Andreelli  , Jardel  , Donner  , Meyer  , Moulin  , Hainque  , Riou  , Laville  , Koivisto  , Vidal
Published: March 2003

Background

An association with subcutaneous adipose tissue TNFα expression and insulin resistance has been suggested in obesity/type‐2 diabetes, but this has not been examined directly. In the first part of the study we investigated whether this association is present in 7 lean, 10 obese nondiabetic and 9 type‐2 diabetic men. In the second part of the study we examined the relationship between adipose tissue TNFα mRNA levels and BMI in 81 nondiabetic subjects spanning a wide range of BMIs.

Methods

Subcutaneous adipose tissue TNFα mRNA levels and insulin sensitivity were determined with quantitative RT‐competitive PCR and hyperinsulinaemic clamp, respectively.

Results.

Subcutaneous adipose tissue TNFα mRNA levels were similar in 7 lean and 10 obese nondiabetic and 9 type‐2 diabetic men (P = 0.68), and did not change in response to 240‐min hyperinsulinaemia. TNFα mRNA levels and insulin sensitivity were not correlated. Unexpectedly, no correlation between TNFα mRNA and BMI was found. The relationship between adipose tissue TNFα mRNA and BMI was examined further in 31 male and 50 female nondiabetic subjects. The subcutaneous adipose tissue TNFα mRNA level correlated with BMI in all subjects (rS = 0.32, P < 0.01), and in a subgroup analysis in men (rS = 0.55, P < 0.01) but not in women (rS = − 0.08). The correlation in men was dependent on a fourfold higher TNFα mRNA level in 5 morbidly obese men while there was no difference in TNFα mRNA levels in lean or obese men.

Conclusions

Subcutaneous adipose tissue TNFα expression does not correlate with insulin sensitivity in nondiabetic or type‐2 diabetic men; is not regulated by acute hyperinsulinaemia; and is increased only in morbidly obese men.
Effects of glucose ingestion on cardiac autonomic nervous system in healthy centenarians: differences with aged subjects
Authors: Paolisso  , Manzella  , Rizzo  , Barbieri  , Gambardella  , Varricchio
Published: March 2003

Background

Spectral analysis of heart rate variability (HRV) investigates the cardiac autonomic nervous system (ANS) activity. In particular, low frequency/high frequency (LF/HF) is considered an index of cardiac sympatho‐vagal balance and is stimulated by glucose ingestion in healthy subjects. No studies have evaluated the effect of glucose ingestion on cardiac ANS in centenarians.

Materials and methods

In 30 healthy centenarians (HC) and 25 aged subjects (AS) power spectral analysis of HRV was investigated during an oral glucose ingestion.

Results

Glucose ingestion rose LF/HF ratio in both groups studied. Such stimulatory effects were restrained to the first 60 min of the study. Independent of age, gender, body mass index (BMI) and fasting plasma norepinephrine and FT3 concentrations, HC had basal total power (1318 ± 546 vs. 1918 ± 818 msec2, P < 0.01), lower low frequency (LF) (33 ± 21 vs. 50 ± 11 n.u. , P < 0.03), and higher high frequency (HF) (74 ± 18 vs. 43 ± 15 n.u., P < 0.05) than AS. Consequently, LF/HF ratio (0.43 ± 0.07 vs. 0.91 ± 0.05, P < 0.02) was also lower in HC than in AS. In AS, but not in HC, the baseline LF/HF ratio correlated significantly with BMI (r = 0.48, P < 0.01), waist‐hip‐ratio (WHR) (r = 0.45, P < 0.02), fasting plasma insulin (r = 0.49, P < 0.01) and norepinephrine (r = 0.57, P < 0.02) concentration. Glucose ingestion was associated with a significant rise in LF/HF ratio in both groups studied but per cent changes in glucose mediated stimulation of LF/HF was lower in HC than in AS. In a control study, water administration did not affect power spectral parameters of HRV.

Conclusion

Our study demonstrates that basal‐ and glucose‐stimulated LF/HF, an indirect index of cardiac sympatho‐vagal balance, are lower in HC than in AS.
Telomerase activity in needle biopsies from prostate cancer and benign prostates
Authors: Wymenga  , Wisman  , Veenstra  , Ruiters  , Mensink
Published: March 2003

Background

Telomerase activation is thought to be essential for the immortality of cancer cells. It may be a prognostic factor in small volume well differentiated prostate cancers and hence a guide for the aggressiveness of the approach. The length of the chromosome tips (telomeres) are maintained by a specific enzyme (telomerase) independently of the normal cell division cycle. Although telomerase is not expressed in most normal human tissues, it is expressed in most human tumours. For the detection of telomerase in small prostate needle biopsy samples a recently developed telomeric repeat amplification protocol (TRAP) assay was used. The aim of the present study was: to measure telomerase activity in human prostate samples, and to evaluate the applicability of this assay on specimens from a prostate biopsy.

Materials and methods

From 36 patients referred for lower urinary tract symptoms (LUTS) or suspicion of having prostate cancer a total of 288 prostate biopsy samples were obtained (8 in each patient). When the digital rectal examination was abnormal and/or when the PSA level was elevated in L.U.T.S., or asymptomatic patients' tissue samples were obtained by transrectal ultrasound (TRUS) guided biopsies. Samples were tested for telomerase activity by a modified TRAP and forwarded for histology.

Results

In 19 out of 36 patients prostate cancer was diagnosed on histology. In 11 of these 19 tumours substantial telomerase activity was detected, whereas only very low telomerase activity existed in 2 of 17 samples from benign prostatic hypertrophy (BPH) patients. In this small series the relative telomerase activity in prostate cancer correlated with histopathological grade.

Conclusions

Our results show the applicability of a TRAP assay to measure telomerase activity in small needle biopsied prostate samples. In poorly differentiated and metastatic cancer we observed that levels of telomerase activity were high. To establish accuracy and to distinguish the ‘relative good from the ugly’ further study is needed.
Serum concentration of 5‐S‐cysteinyldopa in patients with melanoma
Authors: Bánfalvi T. , Gilde K. , Boldizsár M. , Fejös Z. , Horváth B. , Liszkay G. , Beczássy E. , Kremmer T.
Published: March 2003

Background

5‐S‐cysteinyldopa (5‐S‐DC) is a precursor of melanin. Its serum and urinary level can reflect melanoma progression. In this study we examined the concentration changes of 5‐S‐CD in melanomas of different clinical stages and in patients with different symptoms of melanoma, during and after treatment.

Method

Serum samples were taken from 252 melanoma patients on 765 occasions, from June 1996 to July 1998. Levels of 5‐S‐CD were determined by HPLC.

Results

The value of 5‐S‐CD in patients with primary melanoma and in patients without symptoms ranged around the normal level. There was a significant difference between the values of patients with or without symptoms. There was also a significant difference between the 5‐S‐CD values at clinical Stage I and Stage III, as well as at clinical Stage II and Stage III, respectively. Analysing the values of patients with symptoms we found a significant difference between the mean values of primary tumour and stage III, between values in lymph node metastasis and stage III, between values in lung metastasis and stage III. The tumour burden was found to correlate with a rising marker level. In 7% of the symptomatic patients that had a marker level under the upper limit, amelanotic primary tumour was detected.

Conclusion

According to the high marker level in lung and liver metastases, the marker might be useful in monitoring both patients with disease free ocular melanomas, to detect liver metastasis and high‐risk patients after surgical removal of the primary tumour to reveal lung metastases.
Mutations and deletions in core promoter and precore stop codon in relation to viral replication and liver damage in Singaporean hepatitis B virus carriers
Authors: Chen  , Oon
Published: March 2003

Background

The core promoter of hepatitis B virus (HBV) is crucial for the viral replication and mutations may lead to the establishment of chronic infection and development of liver diseases. We analysed this region in Singaporean HBV carriers and assessed their association with viral replication and liver damage.

Materials and Methods

Thirty‐three Singaporean HBV carriers were selected. Serological markers for HBV infection and indicators for liver functions were analysed using commercial kits. Among these patients, 17 were chronic carriers, 10 had cirrhotic livers and 6 others had hepatocellular carcinoma (HCC). The region on the HBV genome covering the entire core promoter and core gene was amplified for each patient by polymerase chain reaction. The amplified DNA fragments were sequenced and analysed.

Results

The incidence of mutations in the core promoter or the precore gene product (stop codon at amino acid 28) was not significantly higher compared with the wild type sequences in patients with liver damage. Most mutations in either the core promoter or precore gene significantly reduced the viral replication, as indicated by HBV DNA levels. High levels of HBV DNA were found in three mutants with deletion in the same region, presumably the binding site of liver enriched factor, within the core promoter.

Conclusion

Our findings revealed a different mutation pattern in the core promoter in Singaporean HBV carriers. While most mutations may not be directly associated with the development of liver diseases, deletions in the core promoter could contribute to enhanced viral replication and should be studied further.
Effects of comicronised fenofibrate on lipid and insulin sensitivity in patients with polymetabolic syndrome X
Authors: Idzior‐Walus B. , Sieradzki J. , Rostworowski W. , Zdzienicka A. , Kawalec E. , Wójcik J. , Żarnecki A. , Blane G.
Published: March 2003

Background

This study investigated the effects of comicronised fenofibrate in patients with dyslipidemia and polymetabolic syndrome X.

Design

After a 6‐week dietary run‐in phase, 37 male patients eligible on lipid criteria entered a 12‐week treatment phase consisting of diet plus one capsule daily containing 200 mg of comicronised fenofibrate (Lipanthyl®).

Results

A significant reduction in plasma concentrations of total cholesterol, LDL cholesterol and triglyceride was observed after 4, 8 and 12 weeks of treatment with fenofibrate. The improvement in the atherogenic index LDL/HDL cholesterol from a pretreatment 3.8 to 3.0 after treatment was highly statistically significant and may be judged as satisfactory. Significant changes were also observed in haemostatic factors (fibrinogen reduced by 19%, factor VII activity reduced by 18%). Fasting serum insulin levels and insulin response (area under the curve) after oral glucose load were significantly reduced by 26.8% and 18.7%, respectively, indicating an improvement of insulin sensitivity. Systolic and diastolic blood pressure were significantly reduced. Uric acid was significantly reduced by 21.6%.

Conclusion

These favourable effects of comicronised fenofibrate both on lipid and non lipid parameters, including insulin sensitivity, may confer to this product a particular interest in the treatment of patients with polymetabolic syndrome X.
Cytokine secretion is impaired in women with diabetes mellitus
Authors: Geerlings S. E. , Brouwer E. C. , Van Kessel K C. P. M. , Gaastra W. , Stolk R P. , Hoepelman A I. M.
Published: March 2003

As women with diabetes mellitus (DM) have an increased prevalence of asymptomatic bacteriuria (ASB) and it is known that a correlation exists between the increased prevalence of genitourinary tract infection and impaired cytokine production in women infected with Human Immunodeficiency Virus (HIV), we studied urinary cytokine excretion in diabetic women and compared it with that of nondiabetic controls.

To evaluate the cytokine secretion capacity of women with DM, both whole blood and isolated monocytes of women with and without DM were stimulated in vitro with lipopolysaccharide (LPS).

Lower urinary interleukin‐8 (IL‐8) and interleukin‐6 (IL‐6) concentrations (P = 0·1 and P < 0·001, respectively) were found in diabetic women than in nondiabetic controls. A lower urinary leukocyte cell count correlated with lower urinary IL‐8 and IL‐6 concentrations (P < 0·05). Lower tumour necrosis factor‐alpha (TNF‐α) and IL‐6, but comparable interleukin‐10 (IL‐10) concentrations were found in whole blood (P < 0·04) and isolated monocytes (P = 0·03) of women with DM type 1 compared to women without DM.

Diabetic women with ASB have lower urinary IL‐6 concentrations than nondiabetic bacteriuric controls. In addition, monocytes of women with DM type 1 secrete lower pro‐inflammatory cytokines after stimulation with LPS than monocytes of women without DM. This is not due to an inhibitory effect of the anti‐inflammatory cytokine IL‐10. This can have important consequences for both host defense, endothelial cell functioning and atherogenesis.
Serum tumour necrosis factor‐α levels in cancer patients are discontinuous and correlate with weight loss
Authors: Bossola M. , Muscaritoli M. , Bellantone R. , Pacelli F. , Cascino A. , Sgadari A. , Battaglia F. , Piccioni E. , Scambia G. , Doglietto G. B. , Fanelli F. Rossi
Published: March 2003

Tumour necrosis factor‐α (TNF) has been regarded as a potential mediator of cancer cachexia. Assessment of TNF circulating levels in cancer patients and their correlation with weight loss has led to controversial results.

We measured TNF circulating levels in 28 patients with gastrointestinal cancer and 29 controls with benign gastrointestinal diseases at different times (08.00 h, 14.00 h, 20.00 h) before operation.

TNF activity was not detected in any of controls at any times. In cancer patients, TNF circulating levels were detectable in 18 cases (64·3%) and appeared to be discontinuous. TNF levels above the limit of detection were present in 15 patients (53·6%) at 08.00 h, in 14 (50%) at 014.00 h and in nine (32·1%) at 20.00 h. Mean TNF levels were 14·3 ± 4 pg mL−1 at 08.00 h, 16·7 ± 4·6 pg mL−1 at 14.00 h (P = 0·05) and 18·5 ± 10·2 pg mL−1 at 20.00 h (P < 0·05 vs. 08.00 h and 14.00 h). According to Spearman's analysis, the sum of TNF concentrations at the three times significantly correlated with the severity of weight loss (Spearman's correlation coefficient =− 0·420; P = 0·026). TNF concentrations were consistently and significantly higher in patients with severe weight loss than in those with moderate or light weight loss at 08.00 h (26·3 ± 8·3, 8·9 ± 4·2, 3·8 ± 2·1, respectively; P = 0·04 at one‐way anova). TNF levels were higher in anorectic than in nonanorectic patients at any hour, but the differences were not statistically significant.

The present study demonstrates that TNF is intermittently or discontinuously detectable in patients with gastrointestinal cancer and that its levels correlate with the severity of weight loss.
The role of interleukin‐6 in the human adrenal gland
Authors: Päth G. , Scherbaum W. A. , Bornstein S. R.
Published: March 2003

Interleukin (IL)‐6 is a potent activator of the hypothalamic–pituitary–adrenal (HPA) axis on all levels in humans, and appears to play a pathogenic role in conditions related to chronic stress and physiological ageing; with physiological ageing showing a similar hormonal and immunological pattern to chronic stress. IL‐6 and its receptor IL‐6R are coexpressed at similar sites in the human adrenal gland, which seems to be an important source of IL‐6 production. In vitro, in primary cultures of adrenal gland cells, chronic exposure to IL‐6 stimulates adrenocortical steroid release in a time‐ and dose‐dependent manner. This explains the high systemic cortisol levels in the absence of adequate plasma concentrations of corticotropin (ACTH) observed in patients after long‐term treatment with IL‐6. It could therefore be concluded that in situations of prolonged stress, when corticotropin‐releasing hormone and ACTH release are suppressed by feedback inhibition due to circulating glucocorticoids, IL‐6 maintains the elevated glucocorticoid levels by direct stimulation of adrenocortical steroidogenesis via autocrine/paracrine mechanisms.
Alterations of lipolytic enzymes and high‐density lipoprotein subfractions induced by physical activity in type 2 diabetes mellitus
Authors: Lehmann R. , Engler H. , Honegger R. , Riesen W. , Spinas G. A.
Published: March 2003

Objective

This study investigates the effects of regular moderate physical activity on lipolytic enzymes and plasma lipid concentration, particularly high‐density lipoprotein cholesterol (HDL‐C) subfractions, in patients with type 2 diabetes mellitus.

Ten patients participated in a 3‐month exercise programme without any changes in current medical therapy. The control group consisted of six patients who were matched with regard to sex, age, diabetes duration and diabetes therapy.

Results

Mean (± SE) physical activity in the intervention group increased from 70 ± 21 to 220 ± 28 min per week, which resulted in an increase in total HDL‐C from 1·04 ± 0·07 to 1·28 ± 0·12 (P < 0·001). The HDL3‐C subfraction increased from 0·71 ± 0·08 to 0·86 ± 0·08 mM (P = 0·01) with no significant changes in the HDL2‐C subfraction (0·33 ± 0·04 vs. 0·42 ± 0·05). These changes were paralleled by an 85% increase in hepatic lipase (HL) activity, from 25·7 ± 5·1 to 47·4 ± 4·9 µmol mL−1 h−1(P < 0·001) and a 45% increase in lipoprotein lipase (LPL) activity, from 16·8 ± 3·0–24·3 ± 2·7 µmol mL−1 h−1(P = 0·01). Lecithin‐cholesterol acyl‐transferase (LCAT) activity increased by 32%, from 156 ± 26 to 206 ± 32 nmol mL−1 h−1(P < 0·001). In the control group there were no significant changes in any of the variables assessed.

Conclusions

Regular moderate physical activity in patients with type 2 diabetes led to an overall increase in HL, LPL, and LCAT. HL showed a more pronounced increase than LPL and LCAT. The changes in lipolytic and transferase enzyme pattern resulted in a significant increase of plasma HDL‐C, mainly of the HDL3‐C subfraction.
Increased levels of biochemical markers of bone turnover in relation to persistent immune activation in common variable immunodeficiency
Authors: Ueland T. , Frøland S S. , Bollerslev J. , Aukrust P.
Published: March 2003

Background Based on the involvement of cytokines and growth factors in bone homeostasis, we hypothesised that patients with common variable immunodeficiency (CVI), characterised by persistent immune activation in vivo, may have disturbed bone metabolism as evaluated by biochemical markers of bone turnover.

Materials and methods Serum levels of tumour necrosis factor α (TNFα), interleukin‐6 (IL‐6), bone‐specific alkaline phosphatase (B‐ALP), osteocalcin, carboxyterminal crosslinking telopeptide of type I collagen (CTX‐I), insulin‐like growth factor (IGF)‐I and IGF binding protein‐3 (IGFBP‐3) were measured in 25 patients with CVI and compared to 25 age‐ and sex‐matched healthy controls.

Results Patients with CVI had significantly higher serum levels of CTX‐I and B‐ALP, and significantly lower serum levels of IGF‐I and IGFBP‐3 compared to controls as shown in cross‐sectional, and as for B‐ALP and CTX‐I, also during longitudinal testing. No differences were observed for osteocalcin between the two groups. The elevated B‐ALP and decreased IGF‐I and IGFBP‐3 levels were most pronounced in a subgroup of CVI patients characterised by persistent activation of proinflammatory cytokines in vivo. Raised B‐ALP and decreased IGF‐I and IGFBP‐3 were also significantly correlated with enhanced IL‐6 and TNF‐α levels in these patients.

Conclusions The present study suggests that persistent immune activation in vivo, with raised levels of proinflammatory cytokines, may be related to disturbed bone homeostasis in CVI patients, further supporting an interaction between immune related mediators and bone metabolism in humans.
Type I collagen markers in cord serum of appropriate vs. small for gestational age infants born during the second half of pregnancy
Authors: Saarela T. , Risteli J. , Kauppila A. , Koivisto M.
Published: March 2003

Background The serum concentration of the N‐terminal propeptide of type I procollagen (PINP) reflects the synthesis rate of type I collagen, whereas the corresponding C‐terminal telopeptide (ICTP) mirrors its degradation.

Design PINP and ICTP were measured in a total of 690 cord serum samples from 592 appropriate‐for‐gestational‐age (AGA) infants and 98 smal‐for‐gestational‐age (SGA) infants. These markers were compared between AGA and SGA infants of different gestational ages, ranging from 23 to 41 weeks, and birth weights, from 620 to 4555 g.

Results Both PINP and ICTP levels were very high in the preterm AGA infants and declined significantly with advancing gestational age, paralleling the shape of the fetal growth velocity curve. Regardless of the quite large interindividual variations observed in these markers, PINP was significantly lower in both the preterm and term AGA infants than in the SGA infants. This was also the case for ICTP in the preterm infants of gestational age less than 36 weeks. In stepwise multiple regression analyses, gestational age, being either AGA or SGA and head circumference were significant factors to explain the levels of PINP and ICTP. The levels of PINP and ICTP were correlated with each other highly significantly in both the AGA and SGA infants (rs = 0·700 and 0·692, respectively; P < 0·001 in both).

Conclusions The levels of type I collagen markers seem to follow closely the shape of the fetal growth velocity curve during different stages of gestation. However, because of the large interindividual variations observed, further studies are needed before the significance of these markers for the assessment of normal and abnormal fetal growth can be established.
Association studies between haemochromatosis gene mutations and the risk of cardiovascular diseases
Authors: Hetet G. , Elbaz A. , Gariepy J. , Nicaud V. , Arveiler D. , Morrison C. , Kee F. , Evans A. , Simon A. , Amarenco P. , Cambien F. , Grandchamp B.
Published: March 2003

Background Haemochromatosis is a common genetic disorder, inherited as an autosomal recessive trait that results in a progressive accumulation of iron in most tissues of the body. Positive association studies have been recently published between cardiovascular diseases and heterozygosity for the major mutation C282Y in the haemochromatosis gene HFE.

Methods In the present work, we have determined the HFE genotypes for C282Y and H63D in subjects from two case‐control studies: the ECTIM and GENIC studies, designed to identify genetic variants associated with myocardial and brain infarction, respectively. In addition, we tested whether HFE mutations were associated with the degree of arteriosclerosis assessed non‐invasively by Doppler ultrasonography on the carotid and femoral arteries, in a group of apparently healthy individuals (the AXA Study).

Results The prevalence of 282Y, and 63D allele carriers, did not differ between cases and controls in the ECTIM and in the GENIC studies, while 63D but not 282Y carriers were more numerous among subjects with atherosclerotic plaques in the AXA Study.

Conclusions These three studies do not provide consistent evidence supporting the hypothesis that HFE mutations are associated with an increased risk of cardiovascular disease and with the development of arteriosclerosis.
Hepatic lipase gene variation is related to coronary reactivity in healthy young men
Authors: Fan Ym , Laaksonen R. , Janatuinen T. , Vesalainen R. , Nuutila P. , Koivula T. , Knuuti J. , Lehtimäki T.
Published: March 2003

Background Impaired coronary flow reserve (CFR) can be used to indicate vascular dysfunction before the appearance of angiographic lesions. The hepatic lipase (HL) gene has a functional promoter polymorphism at position C‐480T, which affects transcription and leads to high activity (C/C) and low activity (C/T, T/T) genotypes. These genotypes modulate HL activity, but their role in coronary artery disease is controversial and the effect on coronary function has not been studied. We investigated whether HL genotypes are associated with coronary artery function in healthy young men.

Materials and methods We studied 49 healthy, mildly hypercholesterolemic men (aged 35 ± 4 years). Myocardial blood flow was measured at rest and during adenosine induced hyperaemia with positron emission tomography using [15O] H2O. HL genotype was determined by PCR and Nla III enzyme digestion.

Results Resting myocardial blood flow was not statistically different in subjects with high and low activity HL genotypes. However, CFR (the ratio of adenosine flow to resting flow) was 24% higher (4·62 ± 1·52 vs. 3·73 ± 1·08 mL g−1 min−1, P = 0·024) in men with the high activity genotype (n = 26) than in those with low activity (n = 23). In multivariate analysis, the HL genotype remained a significant predictor of CFR (P = 0·038) after adjusting for age, body mass index, serum lipids and smoking.

Conclusions The findings of our preliminary study suggest that the C‐480T polymorphism of the HL gene may modify coronary reactivity and reflect differences in the early pathogenesis of coronary dysfunction in these healthy young men. If the association between HL polymorphism and impaired CFR is also present in subjects with other dyslipoproteinemias, the HL polymorphism could be a new risk factor for cardiovascular disease.
Analysis of exocrine pancreatic function in cystic fibrosis: one mild CFTR mutation does not exclude pancreatic insufficiency
Authors: Walkowiak J. , Herzig K.‐H. , Witt M. , Pogorzelski A. , Piotrowski R. , Barra E. , Sobczynska‐Tomaszewska A. , Trawinska‐Bartnicka M. , Strzykala K. , Cichy W. , Sands D. , Rutkiewicz E. , Krawczynski M.
Published: March 2003

Background Cystic fibrosis (CF) is the most common cause of exocrine pancreatic insufficiency in childhood. The aim of the present study is to evaluate the correlation between genotype and exocrine pancreatic insufficiency in CF patients. The special emphasis was put on the analysis of mild CFTR mutations.

Design The study comprised 394 CF patients and 105 healthy subjects (HS). Elastase‐1 concentrations were measured in all subjects.

Results Severe pancreatic insufficiency was associated with the presence of two CFTR gene mutations (ΔF508, N1303K, CFTR dele 2,3 (21kb), G542X, 1717–1G‐A, R533X, W1282X, 621GT, 2183AAG, R560T, 2184insA and ΔI507, G551D, 895T) and mild insufficiency with the presence of at least one mutation (R117H, 3171insC, A155P2, 138insL, 296 + 1G‐A, E92GK, E217G, 2789 + 5G‐A. 3849 + 1kbC‐T/3849 + 1kbC‐T) genotype resulted in high elastase‐1‐values. However, in case of patients with genotype ΔF508/3849 + 10kbC‐T, 1717–1GA/3849 + 10kbC‐T as well as with ΔF508/R334W, both high and low elastase‐1 concentrations were found. Low E1 values were found in a patient with ΔF508/R347P genotype.

Conclusion Patients who carry two ‘severe’ mutations develop pancreatic insufficiency, whereas those who carry at least one ‘mild’ usually remain pancreatic sufficient. However, the presence of one mild mutation does not exclude pancreatic insufficiency.
Ageing impairs insulin‐mediated vasodilatation but not forearm glucose uptake
Authors: Lind L. , Fugmann A. , Millgård J. , Berne C. , Lithell H.
Published: March 2003

Background It is unclear if insulin‐mediated vasodilatation is altered by ageing and if this affects insulin‐mediated glucose uptake.

Material and methods A 2‐h euglycaemic hyperinsulinaemic clamp (56 mU m−2 min−1) was performed in 10 healthy, nonobese elderly men (70–75 years) and 13 young men (23–28 years). Forearm blood flow (FBF) was measured by venous occlusion plethysmography and forearm glucose uptake was calculated by arterial and venous serum glucose determinations in the forearm.

Results Insulin induced an increase in FBF in the younger men (from 3·9 ± 1·1 SD to 5·9 ± 2·2 mL min−1 100 −1mL tissue, P < 0·001), but this insulin‐mediated vasodilatation was completely blunted in the elderly subjects. Glucose extraction during the clamp was significantly higher in the elderly subjects (1·2 ± 0·76 vs. 0·82 ± 0·37 mmol L−1 at 120 min, P < 0·01), resulting in a similar forearm glucose uptake in the two groups. On the other hand, whole‐body glucose uptake was significantly decreased in the elderly subjects (5·3 ± 1·8 vs. 8·0 ± 1·1 mg kg−1 min−1, P < 0·001).

Conclusion The present study showed that the ability of insulin to induce vasodilatation is blunted in the forearm in healthy, nonobese elderly subjects. However, the elderly compensate for this impairment with an increased glucose extraction from arterial blood to maintain an unaltered forearm glucose uptake.
Reassignment of the EPB4·1 gene to 1p36 and assessment of its involvement in neuroblastomas
Authors: Huang S. , Lichtenauer U. D. , Pack S. , Wang C. , Kim A. C. , Lutchman M. , Koch C. A. , Torres‐Cruz J. , Huang S.‐C. , Benz E. J. , Christiansen H. , Dockhorn‐Dworniczak B. , Poremba C. , Vortmeyer A. O. , Chishti A. H. , Zhuang Z.
Published: March 2003

Objectives EPB4·1 has been previously mapped to human chromosome 1p33‐p34.2. In contradiction to this chromosomal location, we have mapped EPB4·1–1p36 by using fluorescence in situ hybridization and radiation hybrid mapping. In neuroblastomas, deletions of the telomeric end of chromosome 1 (1p36) are the most common genetic aberration.

Methods We investigated whether genetic aberrations of EPB4·1 can be detected in some neuroblastomas by analyzing 72 tumours for EPB4·1 mutation, expression, and alternative splicing pattern. Furthermore, EPB4·1 protein from a neuroblastoma cell line was studied for its subcellular localization.

Results Sequence changes could be detected in 14 out of 72 neuroblastomas, including missense, silent, and intronic changes. Duplex RT‐PCR analysis revealed a subset of 11 tumours expressing significantly low levels of EPB4·1. Significant EPB4·1 sequence changes that were detected included an exon 4 G/A missense mutation (amino acid: V/I) that was shown to be associated with absence of wild‐type EPB4·1 expression (3 tumours), an exon 8 G/A missense mutation (V/M) (1 tumour), and an intronic sequence change that was shown to be associated with the presence of an aberrant transcript (1 tumour). Splicing pattern analysis revealed that all EPB4·1 transcripts from tumours exclude exon 3, a splicing pattern for generating the 135 kDa isoform. EPB4·1 cDNA cloned from a neuroblastoma cell line produced a 135‐kDa protein with a cytoplasm/membrane localization.

Conclusions Out of 72 neuroblastomas we have identified 11 tumours with impaired EPB4·1 expression and 5 tumours with significant sequence changes. We also found that the 135 kDa isoform is the main EPB4·1 product in neuroblastoma. EPB4·1 cDNA from a neuroblastoma cell line produced a 135‐kDa protein and displayed a cytoplasm/membrane localization in transfected cells.
Serum amyloid A and high‐density lipoprotein cholesterol: serum markers of inflammation in sarcoidosis and other systemic disorders
Authors: Salazar A. , Pintó X. , Mañá J.
Published: March 2003

Hypocholesterolemia has been observed in several inflammatory diseases such as rheumatoid arthritis, myeloproliferative disorders, systemic lupus erythematosus and sarcoidosis. Serum amyloid A is an acute‐phase reactant that is related to the high‐density lipoprotein cholesterol. This review discusses the relationship between the activation of the cells of the monocyte‐macrophage system, determined by the serum amyloid A levels, and the lipid metabolism, measured as alterations in plasma lipoprotein concentrations. The mechanisms of this association during acute inflammation are also discussed in this review.
Food deprivation increases post‐heparin lipoprotein lipase activity in humans
Authors: Ruge T. , Svensson A. , Eriksson J. W. , Olivecrona T. , Olivecrona G.
Published: March 2003

Objective To study the effect of fasting on lipoprotein lipase (LPL) activity in human post‐heparin plasma, representing the functional pool of LPL.

Design Fourteen healthy volunteers were recruited for the study. The subjects were fasted for 30 h. Activities of LPL and hepatic lipase (HL), and LPL mass, were measured in pre‐ and post‐heparin plasma in the fed and in the fasted states, respectively. For comparison, LPL and HL activities were measured in pre‐ and post‐heparin plasma from fed and 24‐h‐fasted guinea pigs.

Results Fasting caused a significant drop in the levels of serum insulin, triglycerides and glucose in the human subjects. Post‐heparin LPL activity increased from 79 ± 6·4 mU mL−1 in the fed state to 112 ± 10 mU mL−1 in the fasted state (P < 0·01), while LPL mass was 361 ± 29 in the fed state and 383 ± 28 in the fasted state, respectively (P = 0·6). In contrast, fasting of guinea pigs caused an 80% drop in post‐heparin LPL activity. The effect of fasting on human and guinea pig post‐heparin HL activity were moderate and statistically not significant.

Conclusions In animal models such as rats and guinea pigs, post‐heparin LPL activity decreases on fasting, presumably due to down‐regulation of adipose tissue LPL. In humans, fasting caused increased post‐heparin LPL activity.
Hyperprolactinaemia in patients with heart failure: clinical and immunogenetic correlations
Authors: Limas C. J. , Kroupis C. , Haidaroglou A. , Cokkinos D. V.
Published: March 2003

Background Prolactin represents a stimulatory link between the neuroendocrine and immune systems, but its involvement in the neurohumoral adaptations to heart failure (HF) has not been explored.

Methods We prospectively studied 55 patients (45 males, 10 females, age 48 ± 7 years) with NYHA Class II/III HF due either to dilated cardiomyopathy (CMP) (n = 33) or ischemic heart disease (IHD) (n = 22). Serum prolactin levels were determined by radioimmunoassay, soluble interleukin‐2 receptor (sIL‐2R) levels by enzyme‐linked immunoassay and HLA‐DQ genotyping with PCR. Left ventricular ejection fraction (LVEF) and end‐diastolic diameter (LVEDd) were assessed echocardiographically.

Results Hyperprolactinaemia (17·3 ± 4 ng mL−1[Group I] vs. 4·64 ± 2 ng mL−1[Group II], P < 0·01) was found in 14 patients (8 with IHD, 6 with CMP). The distribution of HLA‐DQB1 alleles was compared in the two groups and showed a significant increase in the frequency of *0301 (86% in Group I vs. 45% in Group II, P < 0·05). Histidine at position 30 of the HLA‐DQB1 gene was found in 22% of Group II but in none of Group I patients. Furthermore, there was an inverse correlation between the presence of histidine at position 30 and the levels of serum prolactin. Both sIL‐2R levels, a marker of T‐cell activation, and concanavalin A‐stimulated lymphocyte proliferation were lower in Group I patients (561 ± 106 vs. 804 ± 109 pg mL−1 and 20·8 ± 4 vs. 37·3 ± 5 cpmX103[3H] thymidine, respectively). LVEF was significantly higher (32 ± 5%) and LVEDd smaller (62·0 ± 6 mm) in Group I compared to Group II (25 ± 4% and 68·0 ± 5 mm, respectively, P < 0·01) patients.

Conclusion Hyperprolactinaemia presents in 25% of patients with HF and may reflect decreased activation of T‐lymphocytes associated with relatively preserved LV systolic function which is under immune‐genetic control at the HLA‐DQ locus.
Oxysterols induce interleukin‐1β production in human macrophages
Authors: Rosklint T. , Ohlsson B. G. , Wiklund O. , Norén K. , Hultén L. M.
Published: March 2003

Abstract Background  Oxysterols are biologically active molecules generated during the oxidation of low‐density lipoprotein or formed enzymatically in vivo. In the atherosclerotic plaque newly recruited macrophages may be exposed to oxysterols present in the plaque. How these oxysterols affect the expression and secretion of inflammatory cytokines such as interleukin‐1β (IL‐1β) in macrophages is not known. Therefore the aim of the present study was to investigate how oxysterols regulate the expression and secretion of IL‐1β in human monocyte‐derived macrophages.

Methods  The IL‐1β messenger RNA (mRNA) expression was analysed by reverse transcription‐polymerase chain reaction, and the IL‐1β protein secretion was measured by enzyme‐linked immunosorbent assay.

Results  A significant, dose‐dependent increase in the secretion of IL‐1β was given by 25‐hydroxycholesterol without the addition of lipopolysaccharide (LPS). At a concentration of 2·5 µg mL−1 this increase was similar to that obtained by endotoxin (LPS, 1 µg mL−1). A transient increase in IL‐1β mRNA expression was found in macrophages incubated with 25‐hydroxycholesterol compared with untreated controls. In addition, 25‐hydroxycholesterol dramatically increased the IL‐1β secretion induced by LPS. At a concentration of 5 µg mL−1 of 25‐hydroxycholesterol the LPS‐induced IL‐1β secretion was increased by about 25‐fold. A similar tendency, but not so consistent, was found for 27‐hydroxycholesterol.

Conclusions  Our results show that oxysterols, and 25‐hydroxycholesterol in particular, may modulate the inflammatory response in human macrophages. Consequently the presence of oxysterols in atherosclerotic tissue may dramatically influence the effect of inflammation.
Antitumour and toxic effects on Wistar rats of two new platinum complexes
Authors: Charalabopoulos K. , Karkabounas S. , Ioachim E. , Papalimneou V. , Syrigos K. , Evangelou A. , Agnantis N. , Hadjiliadis N.
Published: March 2003

Background Cancer chemotherapy in humans based on metal complexes started at the clinical level in the late 1970s with the use of cisplatin, which forms intra‐strand cross‐links with DNA.

Methods Two new platinum complexes of cis‐geometry with the amino acids inosine (ino) and l‐alanine (ala), Pt(ino)2Cl2 and cis‐[Pt(NH3)2(ala)](NO3), respectively, were synthesized and pure samples were obtained by means of flash chromatography. These complexes were tested on benzo(a)pyrene‐induced tumours in Wistar rats to detect their antitumour and toxic effects.

Results There was a statistically significant prolongation of the mean survival time of the animals in the two groups tested (272 ± 18 days and 246 ± 26 days, respectively) compared to the control group (195 ± 22 days) (P < 0·001). Toxic effects included a decrease in leucocyte cell count, mild haemolysis, mild haematuria, mild hepatotoxicity, elevated body temperature and hair loss. All of these effects were reversible after drug discontinuation.

Conclusions The two new platinum complexes described here appear to have an effective antitumour activity without severe toxicity when tested on Wistar rats.
Endothelin ETA receptor‐subtype specific antagonism does not mitigate the acute systemic or renal effects of exogenous angiotensin II in humans
Authors: Bayerle‐Eder M. , Langenberger H. , Pleiner J. , Polska E. , Mensik C. , Eichler H.‐G. , Wolzt M. , Schmetterer L.
Published: March 2003

Background Angiotensin II (Ang II) is assumed to play a pathophysiological role in a variety of vascular diseases. Animal studies indicate that these effects are partly attributed to stimulation of endothelin‐1 (ET‐1) release. The aim of the present study was to investigate whether the acute effects of Ang II on systemic and renal haemodynamics in healthy subjects can be influenced by endothelin ETA‐receptor blockade.

Design The study design was balanced, randomized, placebo‐controlled, double blind, two‐way cross‐over, in 10 healthy male subjects.

Methods Subjects received stepwise increasing intravenous doses of Ang II (0·65, 1·25, 2·5, 5 ng kg−1 min−1 for 15 min per dose level) in the presence or absence of BQ‐123 (60 µg min−1), a specific ETA‐receptor antagonist. Renal plasma flow (RPF) and glomerular filtration rate (GFR) were assessed by the para‐aminohippurate and inulin plasma clearance method, respectively. Renal vascular resistance (RVR) was calculated from mean arterial pressure (MAP) and renal plasma flow.

Results Ang II decreased RPF by 34% and GFR by 9% and increased RVR by 94% and MAP by 27% (ANOVA, P < 0·001 vs. baseline, for all parameters). BQ‐123 did not alter these renal and systemic haemodynamic responses to a significant degree. In addition, BQ‐123 had no significant haemodynamic effect under baseline conditions.

Conclusions Short‐term increase of circulating Ang II levels causes systemic and renal pressor effects, which are not mitigated by endothelin ETA‐receptor blockade. This suggests that the pressor response to Ang II cannot be accounted for by the acute release of vasoactive ET‐1.
Higher antioxidant defences in plasma and low density lipoproteins from rugby players
Authors: Evelson P. , Gambino G. , Travacio M. , Jaita G. , Verona J. , Maroncelli C. , Wikinski R. , Llesuy S. , Brites F.
Published: March 2003

Background Even if physical activity constitutes a well‐known antiatherogenic factor, the precise mechanisms underlying this protective effect are not completely clear.

Materials and methods Lipid and antioxidant profiles were evaluated in 15 well‐trained rugby players and 15 sedentary controls. Lipoprotein fractions were separated by sequential ultracentrifugation and α‐tocopherol content was determined in each fraction by high‐performance liquid chromatography. Susceptibility to in vitro oxidation was also measured in intermediate and low density lipoproteins isolated from both groups of subjects as the production of conjugated dienes.

Results Although the sportsmen were not receiving any special diet or vitamin supplementation they showed a slightly improved lipoprotein profile, mainly represented by increased high density lipoprotein‐cholesterol levels (P < 0·05), and an enhanced antioxidant status. The latter was evidenced by an increment in total radical antioxidant potential (P < 0·001), higher ascorbic acid (P < 0·005) and α‐tocopherol (P < 0·05) plasma concentrations, and elevated activities of superoxide dismutase (P < 0·001) and arylesterase (P < 0·01). Moreover, only the fraction of intermediate and low density lipoproteins from rugby players presented higher α‐tocopherol content in comparison with sedentary controls (484 ± 67 vs. 377 ± 123 µg dL−1, respectively; P < 0·01). Nevertheless, the susceptibility to in vitro oxidation of this lipoprotein fraction was not different between both groups.

Conclusions Given that intermediate density and low density lipoproteins represent the most atherogenic fraction, this finding, in combination with the improved lipid and antioxidant status, would add to the link between regular physical activity and protection against cardiovascular disease.
No increase in mortality and morbidity among carriers of the C282Y mutation of the hereditary haemochromatosis gene in the oldest old: the Leiden 85‐plus Study
Authors: Van Aken M. O. , De Craen A. J. M. , Gussekloo J. , Moghaddam P. Hanifi , Vandenbroucke J. P. , Heijmans B. T. , Slagboom P. E. , Westendorp R. G. J.
Published: March 2003

Background The C282Y mutation in the gene for haemochromatosis (HFE) has been associated with various diseases at middle age. However, recent studies indicate that penetrance of the C282Y mutation is low. We explored the association between the C282Y mutation, iron metabolism, and morbidity and mortality in participants of the Leiden 85‐plus.

Study design A cross‐sectional comparison and prospective follow‐up was conducted in two unselected cohorts of 661 and 552 subjects. All subjects were aged 85 years and over. We determined the prevalence of C282Y homozygous and heterozygous subjects, and the association between the C282Y mutation and iron metabolism, all‐cause and specific causes of death.

Results Prevalence of C282Y homozygosity in both cohorts was 0·2% (1/661 and 1/552, respectively) and of C282Y heterozygosity was 12·4% (82/661) and 11·4% (63/552), respectively. These estimates coincide exactly with reported estimates in younger age groups. Median ferritin level was 97 µg L−1 (IQR 39–162) for heterozygous carriers and 89 µg L−1 (IQR 41–157) for noncarriers (P = 0·66). The serum ferritin concentration for one C282Y homozygous subject, a woman aged 86 years at the time of enrollment in 1986, was 392 µg L−1. Cardiovascular morbidity was comparable between the C282Y heterozygous subjects and the noncarriers in both study cohorts. All‐cause and cardiovascular mortality of carriers of the C282Y mutation was similar to that in noncarriers.

Conclusions We found two C282Y homozygous subjects, illustrating that homozygosity can be compatible with survival in very old ages. C282Y heterozygosity was not associated with history of cardiovascular disease morbidity, all cause mortality, cardiovascular mortality, or biochemical phenotype of haemochromatosis at old age.
Captopril and quinapril reduce reactive oxygen species
Authors: Van Der Giet M. , Erinola M. , Zidek W. , Tepel M.
Published: March 2003

Background Angiotensin‐converting enzyme inhibitors may affect reactive oxygen species in humans in vitro and in vivo. In the present study we evaluated whether angiotensin‐converting enzyme inhibitors may affect NAD(P)H oxidase activity.

Materials and methods The production of reactive oxygen species was measured spectrophotometrically in mononuclear leukocytes using the fluorescent dye, dichlorofluorescein diacetate. The effects of quinaprilat, captopril, enalaprilat and lisinopril on phorbol myristate acetate‐induced reactive oxygen species generation were investigated in vitro. The effects of quinaprilat, captopril, enalaprilat and lisinopril on the NAD(P)H oxidase activity of the mononuclear leukocytes were measured photometrically. In addition, reactive oxygen species were measured before and 4 h after oral administration of quinapril.

Results In vitro, the addition of quinaprilat (72 ± 6% of control; mean ± SEM; n= 19; P < 0·001) and captopril (48 ± 2% of control; n= 19; P < 0·001) significantly reduced the phorbol‐12‐myristate‐13‐acetate‐induced reactive oxygen species generation by the mononuclear leukocytes, whereas enalaprilat and lisinopril showed no effect. The effect of captopril on phorbol‐12‐myristate‐13‐acetate‐induced reactive oxygen species generation in vitro was concentration‐dependent. Quinaprilat and captopril significantly inhibited the NAD(P)H oxidase activity. After the oral administration of 10 mg of quinapril the phorbol‐12‐myristate‐13‐acetate‐induced reactive oxygen species generation by the mononuclear leukocytes was significantly decreased from 1981 ± 292% to 988 ± 141% (n = 14; P < 0·01).

Conclusion Quinapril and captopril decrease the production of reactive oxygen species.