bluuebunny/crossref_metadata_2025 huggingface dataset:
Vasorelaxant effects of oestradiols on guinea pigs: a role for gender differences
Authors: Naderali E. K. , Smith S. L. , Doyle P. J. , Williams G.
Published: March 2003

Various studies have shown vasorelaxation properties for 17α‐ and 17β‐oestradiol.

Here, we studied the effects of gender difference as well as oestrous cycle on oestradiol‐induced vasorelaxation in mesenteric arteries from male and female guinea‐pigs and in main uterine arteries from female guinea‐pigs in vitro.

Both 17α‐ and 17β‐oestradiol (0·5–20 µmol L−1) induced concentration‐dependent relaxation of both mesenteric and uterine arteries preconstricted with either noradrenaline (NA; 10 µmol L−1) or KCl (125 mmol L−1) from both day‐7 and day‐15 female guinea‐pigs. 17β‐oestradiol was more potent in relaxing arteries precontracted with NA than those pretreated with KCl, while 17α‐oestradiol was more effective in relaxing those arteries precontracted with KCl. In mesenteric arteries from male animals, 17α‐oestradiol was significantly (P < 0·001) more potent on arteries precontracted with NA than KCl, an effect opposite to that seen on arteries from female animals. However, both 17α‐ and 17β‐oestradiol were more potent in relaxing arteries from male animals compared with their female counterparts.

These data indicate a possible role for gender differences in the vascular effects of oestradiols.
The hormone sensitive lipase gene in familial combined hyperlipidemia and insulin resistance
Authors: Pihlajamäki J. , Valve R. , Karjalainen L. , Karhapää P. , Vauhkonen I. , Laakso M.
Published: March 2003

Backround Insulin resistance in the most common familial dyslipidemia, familial combined hyperlipidemia (FCHL), could be due to variations in the hormone sensitive lipase (HSL) gene.

Materials and methods The coding region of the HSL gene was screened with the single strand conformation polymorphism analysis in probands of 27 FCHL families with 228 members. In addition, the C‐60G promoter substitution of the HSL gene was determined by the restriction fragment length polymorphism analysis in these subjects.

Results No variants in the coding region of the HSL gene were found and the allele frequencies of the C‐60G promoter substitution and the silent variant (G3138A) in the 3′ untranslated region did not differ between 110 control subjects and 27 probands with FCHL. However, in control women the C‐60G substitution was associated with high body mass index [30·6 ± 0·9 kg m−2 (mean ± SD) in subjects with the C/G genotype and 24·8 ± 4·6 in subjects with the C/C genotype, P = 0·012], and in control men with high rates of insulin‐stimulated whole body glucose uptake (70·1 ± 14·7 vs. 56·7 ± 14·2 µmol kg−1 min−1, P = 0·014). In 228 FCHL family members this substitution was associated with high low‐density lipoprotein cholesterol levels in men (4·51 ± 1·12 vs. 5·17 ± 1·28 mmol L−1, P = 0·049), but not in women.

Conclusions The HSL gene is not a major gene for FCHL. However, the − 60G allele of this gene may affect body weight, insulin sensitivity and serum cholesterol levels.
Effects of OPB‐9195, anti‐glycation agent, on experimental diabetic neuropathy
Authors: Wada R. , Nishizawa Y. , Yagihashi N. , Takeuchi M. , Ishikawa Y. , Yasumura K. , Nakano M. , Yagihashi S.
Published: March 2003

Background Nonenzymatic glycation of neural proteins and their end‐products (advanced glycation end‐products, AGE) have been implicated in the pathogenesis of diabetic neuropathy. We need a development of effective ant‐glycation agents for future clinical use.

Materials and methods We examined the effects of OPB‐9195 (OPB), a new inhibitor of glycation, on the peripheral nerve structure and function in diabetic rats. Eight‐week‐old Wistar rats were made diabetic by streptozotocin (40 mg kg−1, i.v.) and OPB (60 mg kg−1 day−1) was given by gavage for 24 weeks. Age‐ and sex‐matched normal Wistar rats were used for comparison.

Results During the experimental period, OPB treatment did not affect the reduced body weight, elevated levels of blood glucose and glycated haemoglobin in diabetic rats. At the end of the experiment, delayed tibial motor nerve conduction velocity was significantly improved (by 60%) in treated diabetic rats, with reduction of serum AGE levels. Expression of immunoreactive AGE in the sciatic nerve was reduced in treated diabetic rats compared with those in untreated rats. Sciatic nerve (Na+,K+)‐ATPase activity was also restored in treated diabetic rats. On the cross‐sectioned sciatic nerves, positive cells with oxidative stress‐related DNA damage, as expressed by 8‐hydroxy‐2′‐deoxyguanosine, were less in the peripheral nerve of treated diabetic rats compared with those of untreated rats.

Conclusion The current study suggested that OPB is beneficial for the reduction of serum AGE and the prevention of diabetic neuropathy.
Relationship between bone mass density and tensile strength of the skin in women
Authors: Piérard G. E. , Piérard‐Franchimont C. , Vanderplaetsen S. , Franchimont N. , Gaspard U. , Malaise M.
Published: March 2003

Background Skin and bone may be affected similarly during the climacteric and during long‐term corticotherapy. Little is known about the correlation between the respective alterations in bone mass density (BMD) and tensile strength of the skin.

Materials and methods A total of 100 women aged 19–88 years, receiving, or not, hormone replenishment therapy or systemic corticosteroids, were enrolled in the study. Tensile strength of the inner forearm skin was measured using the suction method operated in both the steep and progressive‐force application modes. BMD was measured by dual X‐ray absorptiometry at the hip, femoral neck and lumbar spine.

Results Being heterogeneous, the population of volunteers yielded a wide range of BMD and cutaneous tensile strength values. However, significant correlations were found between BMD and tensile skin parameters. In particular, a positive correlation was yielded between the biological elasticity of skin and the BMD of the hip and femoral neck.

Conclusion An overall correlation is shown between skin elasticity on a relatively sun‐protected area and cortico‐trabecular BMD.
Homocysteine in breast cyst fluid
Authors: Tallova J. , Tomandl J. , Bicikova M. , Simickova M.
Published: March 2003

Background Elevated plasma homocysteine concentrations have been reported in a variety of carcinoma, including those of the breast. The risk of breast cancer is higher in patients suffering from gross cystic disease. The breast cyst fluid contains unusual amounts of low‐ and high‐ molecular substances, including steroid hormones and their conjugates. The present study was undertaken to find out the presence of homocysteine in the fluid filling the cysts and have its concentration compared with other thiols, levels of Na+/K+ ratio and steroid hormones.

Materials and methods Fourteen women suffering from gross cystic disease were enrolled in this study. Cystic concentrations of homocysteine (Hcy), cysteine (Cys), cysteinylglycine (Cys‐Gly) and glutathione (GSH) were determined by high performance liquid chromatography, with fluorescence detection; estradiol (E2), progesterone, allopregnanolone and pregnenolone sulfate (PregS) by RIA methods.

Results Mean levels of Hcy, Cys, Cys‐Gly, Na+/K+, E2 and PregS in the fluid filling the breast cysts were significantly higher than the corresponding plasma concentrations. In addition, a negative correlation was found between cystic Hcy and the Na+/K+ ratio (Rs = −0·72, P = 0·003) and positive correlations between cyst Hcy and estradiol (Rs = 0·64, P = 0·018) and Hcy and PregS (Rs = 0·60, P = 0·025).

Conclusion The study provides the first evidence of thiol concentrations in the breast cyst fluid. The finding of a negative correlation between homocysteine and the Na+/K+ ratio support the idea that the homocysteine concentration in breast cysts might be used clinically as a marker for the development of breast cancer disease.
Fibroblast studies documenting a case of peroxisomal 2‐methylacyl‐CoA racemase deficiency: possible link between racemase deficiency and malabsorption and vitamin K deficiency
Authors: Van Veldhoven P. P. , Meyhi E. , Squires R. H. , Fransen M. , Fournier B. , Brys V. , Bennett M. J. , Mannaerts G. P.
Published: March 2003

Background 2‐Methylacyl‐CoA racemase interconverts the 2‐methyl group of pristanoyl‐CoA or the 25‐methyl group of hydroxylated cholestanoyl‐CoAs, allowing further peroxisomal desaturation of these compounds in man by the branched chain acyl‐CoA oxidase, which recognise only the S‐isomers. Hence, oxidation studies in fibroblasts, currently based on the use of racemic substrates such as [1–14C] pristanic acid, do not allow us to distinguish between a deficient racemase or an impaired oxidase.

Design To evaluate the racemase activity directly, the 2R‐isomer of[1–14C] pristanic acid, as well as the 2R‐isomer of 2‐methyl‐[1–14C] hexadecanoic, a synthetic pristanic acid substitute, were prepared and their degradation by cultured human skin fibroblasts was compared to that of the racemic substrates.

Results In fibroblasts in a young girl, presenting with elevated urinary levels of trihydroxycholestanoic acid metabolites but normal plasma levels of very long chain fatty acids, a partial deficient degradation of racemic [1–14C] pristanic acid was observed. Incorporation of 2R‐[1–14C] pristanic acid in glycerolipids of the patient's fibroblasts proceeded normally, but breakdown was impaired. Similar findings were seen with the 2R‐isomer of 2‐methyl‐[1–14C] hexadecanoic. These data, combined with the fact that the branched chain acyl‐CoA oxidase, catalyzing the first oxidation step of pristanic acid and bile acid intermediates in man, appeared normal, suggested a peroxisomal β‐oxidation defect in the patient at the level of 2‐methylacyl‐CoA racemase.

Conclusion Carboxy‐labelled 2R‐methyl branched chain fatty acids might be useful tools to document cases of racemase deficiencies. Because a brother of the patient died with a diagnosis of vitamin K deficiency, an impaired racemase might be responsible for other cases of unexplicable malabsorption.
Brachial artery pulse pressure and common carotid artery diameter: mutually independent associations with mortality in subjects with a recent history of impaired glucose tolerance
Authors: Van Dijk R. A. J. M. , Dekker J. M. , Nijpels G. , Heine R. J. , Bouter L. M. , Stehouwer C. D. A.
Published: March 2003

Background Decreased large artery function, as reflected by increased brachial artery pulse pressure and increased carotid artery diameter and stiffness, may contribute to the increased mortality risk that is observed in subjects with impaired glucose tolerance. We therefore investigated the association between brachial artery pulse pressure and carotid artery diameter and stiffness, which are estimates of central artery stiffness and arterial remodelling, respectively, and mortality in subjects with a recent history of impaired glucose tolerance.

Design A prospective, population‐based cohort study. We measured brachial artery pulse pressure by oscillometric blood pressure measurements, and common carotid artery diameter and distensibility and compliance coefficients by ultrasound in 140 subjects with a recent history of impaired glucose tolerance. During a median 6·6‐year follow‐up, 16 subjects died.

Results Brachial artery pulse pressure and common carotid artery diameter were positively related to all‐cause mortality [hazard ratios per standard deviation, 1·7 (1·2–2·5) and 2·1 (1·3–3·3), respectively]. Results were similar after adjustment for gender, age, waist‐to‐hip ratio, body mass index, total cholesterol concentration, pre‐existent cardiovascular disease, and hypertension, and after additional mutual adjustment. Common carotid artery distensibility and compliance coefficients were not statistically significantly associated with mortality.

Conclusions Among subjects with a recent history of impaired glucose tolerance, brachial artery pulse pressure and common carotid artery diameter are independently associated with mortality risk. Stiffness of the central arteries may explain the association between pulse pressure and mortality risk. The association between carotid diameter and mortality risk is more likely to reflect arterial remodelling in response to atherosclerosis than that in response to increased local stiffness.
Quantitative, phenotypic, and functional evaluation of basophils in myelodysplastic syndromes
Authors: Füreder W. , Schernthaner G.‐H. , Ghannadan M. , Hauswirth A. , Sperr W. R. , Semper H. , Majlesi Y. , Zwirner J. , Götze O. , Bühring H.‐J. , Lechner K. , Valent P.
Published: March 2003

Background The myelodysplastic syndromes (MDS) are a group of clonal haematological disorders characterized by cytopenia(s), reduced differentiation‐capacity of myeloid cells, and impaired leukocyte function. However, little is known so far about basophil granulocytes in MDS.

Design We have compared the numbers, phenotype and function of basophils in MDS patients with those in healthy subjects. A total numer of 23 patients with MDS (refractory anaemia, n = 8; refractory anaemia with ringsideroblasts, n = 7; refractory anaemia with excess of blasts/refractory anaemia with excess of blasts in transformation, n = 8) and 20 healthy donors were included.

Results The numbers of blood basophils in MDS patients (34·6 ± 62·9 μL−1) was lower compared to healthy controls (58·6 ± 64·9 μL−1). Correspondingly, whole blood histamine levels were lower in MDS patients (MDS 34·1 ± 29·1 ng mL−1 vs. normal donors 72·0 ± 36·9 ng mL−1). Like ‘normal’ basophils, basophils in MDS expressed interleukin‐3 receptor α (CD123), E‐NPP3 (CD203c), CR1 (CD35), CR3 (CD11b), CR4 (CD11c), membrane co‐factor protein (CD46), decay‐accelerating factor (CD55) and membrane attack complex inhibitory factor (CD59), as well as receptors for C3a, C5a (CD88), and IgE. Recombinant human (rh) C5a and anti‐IgE induced significant release of histamine from basophils in both groups of donors without significant differences between MDS and healthy controls.

Conclusions The absolute numbers of basophils in MDS patients are lower than in normal donors. However, basophils in MDS do not differ from their ‘normal counterparts’ in terms of complement receptor expression, IgE‐receptor expression, or functional responses to respective ligands.
Expression of hepatocyte growth factor, keratinocyte growth factor and their receptors in experimental chronic pancreatitis
Authors: Otte J‐M. , Schwenger M. , Brunke G. , Sparmann G. , Emmrich J. , Schmitz F. , Fölsch U. R. , Herzig K.‐H.
Published: March 2003

Background Hepatocyte (HGF) and Keratinocyte growth factors (KGF) are key factors of tissue organization and regeneration. These peptide growth factors and their receptors c‐met and keratinocyte growth factor receptor (KGFR) are overexpressed in pancreatic cancer.

Aim Expression and localization of ligands and receptors were investigated during the development of experimental chronic pancreatitis.

Methods Chronic pancreatitis was induced in rats by intravenous injection of dibutyltin dichloride. One to 60 days after treatment, the expression of growth factors and receptors was analysed by competitive polymerase chain reaction, Western blot analysis and immunohistochemistry.

Results HGF mRNA expression increased (10‐fold) until days 7–14 followed by a decrease to control level. Expression of c‐met mRNA constantly increased (15‐fold). KGF and KGFR mRNA expression were increased after 14–28 days (5‐fold) and then returned to control levels. mRNA expression patterns correlated with changes in the protein expression, whereas protein levels of KGF remained unchanged. Ligands were localized in mesenchymal cells and their receptors on epithelial cells.

Conclusions The significant increase of HGF and c‐met expression suggests an essential role of this growth factor in the morphological changes during the development of chronic pancreatitis. Changes in the expression of KGF and KGFR are less pronounced.
Re‐adaptation of the gastroduodenal mucosa to DNA synthesis during protracted stress
Authors: Rubio C. A. , Sveander M. , Befrits R.
Published: March 2003

Background Previous study showed that under the influence of protracted physical stress, the DNA synthesis (DS) of the gastroduodenal mucosa rises, the highest peak being reached at 4 weeks. At 8 stress weeks, the DS decreases to values similar to those recorded at the beginning of the experiment. The possibility that this DS adaptation (DSA) could be maintained beyond 8 weeks in animals allowed to subsequent resting (stress‐free) weeks, was now explored.

Methods Sixty‐five rats were investigated. Sixty rats were transported to the stress laboratory. Thirty were water plunged and 30 sham handled once a day, 5 days a week, for 8 weeks. After 8 stress weeks, groups of 5 animals were allowed to rest for one, two, three and four weeks, ending with single water plunging or sham handling. All 65 animals received ip injection of 3H‐thymidine before they were sacrificed. The ratio radioactive DNA/total DNA reflected the DS of the stomach and duodenum.

Results DSA was achieved at 8 stress weeks (stomach). The DS in the stomach of stressed rats had significantly increased at 10 weeks (P < 0·05), but at 12 weeks it had decreased to 8 weeks values. The DS in the duodenum of stressed rats had significantly increased at 9, 10 and 11 weeks (P < 0·05), but at 12 weeks DS values had decreased to those of 8 weeks.

Conclusions The DSA at 8 stress weeks had not prevailed because the DS increased considerably in the following weeks. At 9–11 weeks, DS values had significantly increased (P < 0·05) in stressed rats, but at 12 weeks a DS re‐adaptation (DSRA) had occurred. Thus, it took 8 weeks to achieve DSA (stomach), but only 4 weeks to accomplish DSRA. Animals experienced sham handling as an stressor (milder) as it also induced DS re‐adjustments in the gastroduodenal mucosa. Autoradiography showed that the labelling in DNA synthesizing cells was limited almost exclusively to the mucosal layer. The model described may prove of value to studies aimed at abating the disparate fluctuations of DNA synthesis in the gastroduodenal mucosa during the various phases of protracted stress.
Helicobacter pylori masks differences in homocysteine plasma levels between controls and type 2 diabetic patients
Authors: Cenerelli S. , Bonazzi P. , Galeazzi R. , Testa I. , Bonfigli A. R. , Sirolla C. , Giunta S. , Galeazzi L. , Fumelli D. , Testa R.
Published: March 2003

Background Data in the literature have not clarified whether type 2 diabetes mellitus affects homocysteine plasma levels. Different variables able to influence homocysteine could be the cause of these controversial findings. An important but neglected confounding factor is Helicobacter pylori, which has been demonstrated to be a cause of elevated levels of homocysteine and which is prevalent in the Caucasian population, ranging from 30 to 40% incidence. Starting from these findings we wanted to verify whether differences in homocysteine levels exist between a type 2 diabetic population and a control group, taking into account the presence/absence of Helicobacter pylori.

Design The study was carried out on a group of uncomplicated and normotensive type 2 diabetic patients (n = 30, 55·7 ± 9·7 years) and on a control group (n = 43, 51·2 ± 11·3 years). On these subjects we evaluated: main parameters of glyco‐ and lipo‐metabolic balance, presence of Helicobacter pylori by 13C Urea Breath Test, plasma homocysteine, vitamin B12, folate and genetic polymorphism of methylenetetrahydrofolate reductase.

Results Evaluating the two groups as a whole, significant differences in homocysteine were found when considering Helicobacter pylori presence/absence (14·0 ± 6·5 vs. 10·6 ± 4·7 µmol L−1, respectively, P < 0·01) without differences of vitamins and the genetic polymorphism of methylenetetrahydrofolate reductase. The positive interaction found among Helicobacter pylori, diabetes and homocysteine (P = 0·03) taking into account all the other evaluated confounding factors, demonstrates that a significant difference in homocysteine plasma levels exists between diabetics and controls (Helicobacter pylori‐negative: diabetics 12·5 ± 5·6 µmol L−1, controls 9·4 ± 3·8 µmol L−1; Helicobacter pylori‐positive: diabetics 13·6 ± 5·8 µmol L−1, controls 14·3 ± 7·0 µmol L−1).

Conclusions Type 2 diabetes seems to induce per se higher levels of homocysteine, which appears to be one of the factors responsible for the increased risk of vascular damage.
Complement activating antibodies against the human 60 kDa heat shock protein as a new independent family risk factor of coronary heart disease
Authors: Veres A. , Szamosi T. , Ablonczy M. , Szamosi T. , Singh M. , Karádi I. , Romics , L. , Füst G. , Prohászka Z.
Published: March 2003

Background Several groups have reported high levels of antibodies against 60 kDa heat shock proteins (hsp) associated with coronary heart disease.

Methods and results Complement activating (CA) antihsp60 autoantibodies were measured by the AtheroRisk kit (CardioPath Ltd, Alloa, UK), in parallel with IgG antibodies to human hsp60 and mycobacterial hsp65 by ELISA in 32 healthy children (18 boys, 14 girls, 11·8 ± 4·0 years). At least one of the parents of these children had a history of myocardial infarction before 55 years of age (high family risk (HFR) group). The control group consisted of 63 healthy children (31 boys, 32 girls, 9·0 ± 3·6 years) without known family history of coronary heart disease (CHD), hypertension, and diabetes mellitus. Concentrations of CA antihsp60 antibodies were significantly (P = 0·021) higher in the HFR group than in the control group. Also in the HFR group, significantly (P = 0·004) lower high‐density lipoprotein cholesterol (HDL‐C)‐cholesterol (measured enzymatically) and significantly (P = 0·020) higher low‐density lipoprotein cholesterol (LDL‐C)‐cholesterol levels (calculated by the Friedewald formula) were observed when compared with the controls. The difference in the CA antihsp60 antibody levels between the HFR and control groups remained significant even after adjustments for age, smoking, HDL‐cholesterol, LDL‐cholesterol levels, and white blood cell count. Children with high (in the highest quartile) CA antihsp60 antibody levels compared with those with normal levels of these antibodies also had adjusted odds ratios (OR) of 9·80 (2·15–44·58, P = 0·003), indicating high family risk. No significant difference in the IgG antihsp antibody levels was observed.

Conclusions These findings indicate that high levels of CA autoantibodies against hsp60 can be considered to be a novel family risk factor of CHD, independent of HDL‐ and LDL‐cholesterol levels.
Effects of hormone replacement therapy on blood platelets
Authors: Thijs A. , Van Baal W. M. , Van Der Mooren M. J. , Kenemans P. , Dräger A. M. , Huijgens P. C. , Stehouwer C. D. A.
Published: March 2003

Background Hormone replacement therapy (HRT) increases the risk of cardiovascular morbidity in postmenopausal women under certain circumstances. Part of this effect may be the result of the influence of HRT on blood platelets. We studied the effect of short‐term oral hormone replacement therapy (unopposed oestradiol or sequentially combined oestradiol and trimegestone or dydrogesterone) on platelet activation parameters in healthy postmenopausal women.

Design We designed a prospective, randomised, placebo‐controlled 12‐week study. Sixty healthy, normotensive, nonhysterectomised, postmenopausal women received daily micronised oestradiol (E2) 2 mg (n = 16), or 2 mg E2 daily sequentially combined with either trimegestone 0·5 mg daily (n = 14) or dydrogesterone 10 mg daily (n = 14), or placebo (n = 16). Data on platelet activation were collected at baseline and after 12 weeks of treatment using flow cytometry.

Results Twelve weeks of treatment with combined HRT was associated with an increase in platelet activation parameters P‐selectin and glycoprotein 53 (by 17% and 14%, respectively, P = 0·04 vs. the placebo group for both comparisons), suggesting alpha granule and lysosome degranulation. E2 replacement therapy was associated with an increase in P‐selectin labelling by 22% (P = 0·04 vs. the placebo group).

Conclusion Short‐term treatment with oestradiol or combined HRT increases the amount of circulating activated platelets as measured by flow cytometry. This could be a mechanism by which short‐term HRT might increase the risk of thrombosis.
Effect of nicotine replacement and quitting smoking on circulating adhesion molecule profiles (sICAM‐1, sCD44v5, sCD44v6)
Authors: Palmer R. M. , Stapleton J. A. , Sutherland G. , Coward P. Y. , Wilson R. F. , Scott D. A.
Published: March 2003

Background Soluble ICAM‐1 (sICAM‐1; sCD54), sCD44v5 and sCD44v6 are circulating adhesion molecules, with immunomodulatory potential, that have been frequently attributed diagnostic, prognostic and aetiological significance in a number of inflammatory and malignant diseases. We have previously shown that systemic concentrations of these molecules are increased significantly in tobacco smokers, but reduce to within normal levels at 12 months following successful quitting.

Materials and methods We have been able to extend these observations by measuring levels before and 4, 8, 22 and 52 weeks after smoking cessation in subjects receiving high‐dose nicotine replacement therapy (25 mg of nicotine; n= 34) or placebo patches (n = 34) for 26 weeks. Smoking cessation was confirmed by regular measurement of expired‐air CO levels and by plasma cotinine analysis.

Results Plasma sICAM‐1, sCD44v5 and sCD44v6 concentrations all declined rapidly within 4 weeks of smoking cessation (P < 0·001 for all declines). Additionally, no differences were observed between those using nicotine replacement and those who were not for sICAM‐1, sCD44v5, or sCD44v6.

Conclusions The recovery in smoking‐associated adhesion molecule profiles represents an almost immediate beneficial effect of smoking cessation. Nicotine replacement therapy is an effective aid to quitting and does not affect these recoveries. The elevated levels of these important risk factors in smokers (sICAM‐1, sCD44v5 and sCD44v6) are linked to noxious element(s) in tobacco smoke other than nicotine or nicotine metabolites.
Red blood cell antioxidant and iron status in alcoholic and nonalcoholic cirrhosis
Authors: Fiorelli G. , De Feo T. M. , Duca L. , Tavazzi D. , Nava I. , Fargion S. , Cappellini M. D.
Published: March 2003

Background Iron overload has been reported in alcoholic liver cirrhosis but it remains to be established whether iron is involved in inducing oxidative damage to erythrocytes in alcoholic cirrhosis. The aim of this study was to assess oxidative damage and red cell indicators of antioxidant defences in alcoholics with mild‐to‐severe liver cirrhosis, taking into account the iron status.

Materials and methods Twenty‐nine patients with alcoholic liver cirrhosis (AC) and 27 with nonalcoholic cirrhosis (NAC) were studied. Serum lipid peroxides (LPO) were assayed by a colourimetric method. Serum‐free malonyldialdehyde (MDA) was assayed by selected ion monitoring in positive chemical ionization; serum 4‐hydroxy‐2(E)‐nonenal (4‐HNE) was determined by a colorimetric method. Reduced (GSH) and oxidized glutathione (GSSG), adenine and pyridine cofactors were assayed in whole blood extracts by HPLC. Hexose‐monophosphate shunt (HMPS), glycolytic pathway (EMP) and antioxidant enzyme activities were determined by standard methods. Iron status was evaluated by standard clinical chemistry and by histological grading of liver iron. Nontransferrin‐bound iron (NTBI) was measured in serum by HPLC.

Results GSH progressively decreased with increasing severity of liver involvement in AC and NAC. MDA, 4‐HNE and NTBI were significantly higher in AC serum. Stimulation of red cell HMPS and reducing potential, in terms of NADPH production, were more pronounced in AC.

Conclusions

These results suggest that NTBI is more important than the decrease of antioxidant defences in inducing lipid peroxidation. NTBI may play a catalytic role in free radical reactions in the presence of cellular reductants such as NADPH.
Inhibition of endogenous pancreatic enzyme secretion by oral pancreatic enzyme treatment
Authors: Walkowiak J. , Witmanowski H. , Strzykala K. , Bychowiec B. , Songin T. , Borski K. , Herzig K.‐H.
Published: March 2003

Background  The existence of a feedback mechanism for exocrine pancreatic secretion in humans is controversial. Exclusion of proteases from the duodenum stimulates exocrine pancreatic secretion. Conversely, addition of exogenous enzymes could reduce the enzyme secretion. Further investigation of the feedback mechanism should be performed under the most physiological conditions. In the present study we investigated exocrine pancreatic function by measuring fecal enzyme output in healthy volunteers consuming a normal diet, before and during a time course of exogenous pancreatic enzyme supplementation.

Material and methods  Twenty‐five healthy subjects (HS) were given two different doses (30 and 60 FIP proteases kg−1 d−1) divided by the number of meals. In all subjects, fecal elastase‐1 (E1) concentrations and chymotrypsin (ChT) activities were measured without and with enzyme supplements after 7 days of treatment. In eight subjects, E1 concentrations and ChT activities were measured daily for 10 consecutive days. The subjects were given a dose regimen of 100 FIP proteases kg−1 d−1 (divided by the number of meals) for the first 7 days.

Results  Oral pancreatic treatment dose‐dependently inhibited endogenous pancreatic secretion measured with the use of E1 concentrations. In both regimen groups, the differences were statistically significant. The exogenous enzymes, which interfere with colorimetric method for ChT, dose‐dependently increased ChT output. However, only the higher dose resulted in a statistically significant difference. In the subgroup of eight HS, time‐dependent changes of fecal enzyme output occurred with a decrease of E1 concentrations and an increase of ChT activity from the second up to eighth or ninth day of the experiment.

Conclusion  Exogenous applied pancreatic enzymes, dose‐ and time‐dependently inhibited endogenous pancreatic secretion. The obtained results strongly support the existence of a protease mediated feedback mechanism in humans.
Polymorphism of Old Kladruber horses, a surviving but endangered baroque breed
Authors: Horín P. , Cothran E. G. , Trtková K. , Marti E. , Glasnák V. , Henney P. , Vyskocil M. , Lazary S.
Published: March 2003

Analysis of MHC class I and class II polymorphism, as well as data from other polymorphic systems (non‐MHC lymphocyte alloantigen, blood groups systems, biochemical polymorphisms and microsatellite loci), was used to characterize the extent and distribution of the genic polymorphism of Kladruber horses. A breed‐characteristic distribution of the MHC polymorphism was found. The repertoire of defined MHC class I specificities was restricted, especially in the grey subpopulation and in stallions, but a high frequency of blanks suggests the possible existence of undetected specificities. Despite the small population size and a relatively high degree of inbreeding, high heterozygosity in MHC haplotypes has been conserved. The extent of polymorphism and the degree of heterozygosity in other loci were also relatively high. A comparison of the two existing subpopulations, grey and black, at all the loci tested, including RAPD markers, characterized them as genetically distinct, although clearly related. The genetic distances between them were of the same order of magnitude as between distinct breeds. The results may be useful in defining short‐term and long‐term breeding policy within the breed and for further studies of associations with disease and other traits.
Phylogenetic analysis of primate MIC (PERB11) sequences suggests that the representation of the gene family differs in different primates: comparison of MIC (PERB11) and C4
Authors: Cattley S. K. , Longman N. , Dawkins R. L. , Gaudieri S. , Kulski J. K. , Leelayuwat C.
Published: March 2003

Duplication of segments within the MHC has led to numerous multicopy families such as class I, class II, C4 and MIC (PERB11). Different copy numbers between haplotypes and species may be explained by the extent of duplication and subsequent deletion. There are at least five copies of MIC (PERB11) in humans, but MICA (PERB11.1) appears to have been deleted from the chimpanzee.

By comparing the sequences of primates (chimpanzee, gorilla, gibbon, orang‐utan, pygmy chimpanzee, Patas monkey, Aethiops and baboon) we conclude that the gorilla has a copy of PERB11.1, whereas the baboon and Patas possess MICD (PERB11.4) and/or MICE (PERB11.5) rather than MICA (PERB11.1). These findings indicate that the primate MHC is more plastic than has been appreciated.
Inverted ratio of m‐fas/s‐fas expression in early onset pauciarticular juvenile chronic arthritis
Authors: Haas J.‐P. , Frank C. , Haefner R. , Herrmann M. , Späth H. , Ruder H.
Published: March 2003

The expression of m‐fas/apo1 (CD95) and its soluble form (s‐fas) was studied in patients with early onset pauciarticular (n = 23) and systemic juvenile chronic arthritis (n = 7). RNA was prepared from peripheral blood of patients and 22 healthy controls and from 23 samples derived from the synovial fluid of JCA patients. The ratio of m‐fas/s‐fas transcripts was calculated by densitometry of fas‐specific RT‐PCR products. An inverted ratio of m‐fas/s‐fas transcripts was found in PBMC and mononuclear cells from the joints of early onset pauciarticular JCA patients. The m‐fas/s‐fas ratios were similar in PBMC and mononuclear cells from synovial fluid. PBMC from patients with a systemic JCA showed the same m‐fas/s‐fas ratio as healthy controls, with an inverted ratio of both transcripts in cells from the synovial fluid. The data indicate a role of Fas/Apo1 regulated apoptosis in EOPA‐JCA which is not limited to the affected joints.
Human platelet antigens: typing by PCR using sequence‐specific primers and their distribution in blood donors resident in Wales
Authors: Sellers J. , Thompson J. , Guttridge M. G. , Darke C.
Published: March 2003

We developed and validated HPA‐1 to HPA‐6 typing by PCR–SSP using a combination of established, modified and newly designed sequence‐specific primers. We confirmed that the PCR primer mixtures functioned under the same PCR conditions as our standard HLA‐A, ‐B, ‐C, ‐DR, ‐DQ PCR–SSP typing system. This allows concurrent testing for both HPA and HLA specificities and is therefore the system of choice for both clinical and large‐scale blood donor panel HPA and HLA typing by PCR–SSP. Test validation included typing a population of blood donors living in Wales. These HPA frequencies were consistent with those of other European Caucasoid populations. HPA‐4b and ‐6b were absent and HPA‐5b, which shows some frequency variation, had a phenotype frequency of 18.9% (allele frequency 0.0973), being close to that of the Dutch (19.7%) and Austrian (20.4%) populations and almost twice that found in Finns (10.0%). HPA genotype frequencies showed a good fit to Hardy–Weinberg equilibrium, further supporting the validity of our typing method.
Rare polymorphisms in the promoter regions of the human interleukin‐12 p35 and interleukin‐12 p40 subunit genes
Authors: Pravica Vera , Brogan Iain J. , Hutchinson Ian V.
Published: March 2003

We have described two rare, single‐nucleotide polymorphisms within the promoter regions of the IL‐12 p35 and p40 subunit genes. Transitions C to T at position −916 from the transcription start of the IL‐12 p35 gene and G to T at position −1287 in the IL‐12 p40 gene were observed.
Molecular cloning and sequencing of a cDNA encoding the feline cytotoxic T‐lymphocyte‐associated protein 4 (CTLA4) homologue *
Authors: Nishimura Y. , Miyazawa T. , Ikeda Y. , Izumiya Y. , Nakamura K. , Sato E. , Mikami T. , Takahashi E.
Published: March 2003

The feline CTLA4 cDNA encodes a transmembrane protein which shares 87.9% sequence identity with the human CTLA4 molecule. The cytoplasmic region of the feline CTLA4 is identical to that of humans and mice, which suggests conserved function(s) such as the regulation of cell‐surface expression of this molecule.
A polymorphic major histocompatibility complex class II‐like locus maps outside of both the chicken B‐system and Rfp‐Y‐system
Authors: Juul‐Madsen H. R. , Dalgaard T. S. , Guldbrandtsen B. , Salomonsen J.
Published: March 2003

Chickens have two major regions encoding major histocompatibility complex (MHC) class Iα genes and MHC class IIß genes, the serological and functional B‐system and the Rfp‐Y‐system. Recently, they have been shown to assort in a genetically independent way although still located on the same microchromosome. Moreover, the monomorphic MHC class IIα gene maps at a third locus located 5 c m from the nearest class IIß genes, located in the B‐system (
Kaufman et al., 1995
). A pedigree family was studied in three generations in order to assign MHC class IIß restriction fragments observed in Southern blot analyses to either the B‐system, the Rfp‐Y‐system or the B‐Lα locus. In this study, we demonstrate by classical genetic testing of chickens within this fully pedigreed family the existence of an MHC class II‐like polymorphic restriction fragment that segregates independently of the B‐system, the Rfp‐Y‐system and of the B‐Lα locus.
Prevalence of CCR2–64I, SDF1–3′A and CCR5‐Δ32 alleles in healthy Thais
Authors: Nookhai S. , Ruxrungtham K. , Phanuphak P. , Oelrichs Robert
Published: March 2003

A genetic survey was performed of 200 healthy Thai blood donors for the frequency of three alleles that influence susceptibility to HIV infection and the rate of progression to HIV disease. The CCR5‐Δ32 allele was not detected in this population. The CCR2–64I allele was detected at a frequency similar to that found in other Asian populations (15.7%). SDF1–3′A was detected at 33.2%, supporting a cline of increasing frequency of this allele from African and Caucasian to Asian (particularly Australasian) populations. These results have implications for the role of host genetic background in the biology and pathology of HIV in Thailand, and indicate that a systematic survey of non‐Caucasian populations may reveal novel alleles important in HIV disease.
New chimpanzee MHC class I alleles cloned by polymerase chain reaction
Authors: Machida S. , Matsui M. , Tanoue T. , Hayasaka I. , Akatsuka T.
Published: March 2003

We describe polymerase chain reaction (PCR) cloning of full‐length Patr‐A, ‐B and ‐Cw locus alleles from a chimpanzee using one sense/antisense primer pair. Of the six alleles cloned here, two have not previously been described. Comparison of nucleotide sequences between the two novel Patr alleles and various HLA alleles suggests that the new Patr‐A allele belongs to the A1/A3/A11 family, whereas the new Patr‐Cw allele is not closely related to any known HLA‐Cw allele.
Organization of the mouse macrophage C‐type lectin (Mcl) gene and identification of a subgroup of related lectin molecules
Authors: Balch S. G. , Greaves D. R. , Gordon S. , McKnight A. J.
Published: March 2003

A number of genes encoding C‐type lectin molecules have been mapped to the natural killer gene complex (NKC) at the distal region of mouse chromosome 6 and to a syntenic region on human chromosome 12p12‐p13. In addition to those receptors which regulate NK cell function, related structures expressed on other cells types have also been localized to this chromosomal region. Among these are a number of recently characterized genes, including macrophage C‐type lectin (MCL), macrophage‐inducible C‐type lectin (Mincle), dendritic cell immunoreceptor (DCIR) and dendritic cell‐associated lectin‐2 (Dectin‐2). The amino acid sequences comprising the single C‐type lectin domains of MCL, Mincle, DCIR and Dectin‐2 are shown here to be closely related to each other. These molecules show overall similarity to two groups of animal C‐type lectins, groups II and V, which demonstrate type II transmembrane topology. In this study, sequence analysis suggests that MCL, Mincle, DCIR and Dectin‐2 represent a subset of group II‐related C‐type lectins which may participate in analogous recognition events on macrophages and dendritic cells. The genomic organization of the MCL gene and the sequence of the promoter region, with putative regulatory elements, were determined from a mouse MCL genomic DNA clone and are described here in detail.
Beta‐2 microglobulin‐free HLA class I heavy chain (FHC) A3 and/or A30 soluble products contribute only minimally to serum FHC expression
Authors: Perosa F. , Prete M. , Luccarelli G. , Favoino B. , Dammacco F.
Published: March 2003

No monoclonal antibodies (mAbs) are presently available to measure the total amount of β2‐microglobulin‐free HLA class I heavy chain (FHC) in sera. The available ELISA‐based double determinant immunoassay (DDIA), established to measure FHC, uses two mAbs (TP25.99 and HC‐10) that recognize a monomorphic determinant expressed on all HLA‐B/C FHC products and a determinant expressed only on some HLA‐A FHC products. This restricted reactivity implies that, in addition to HLA‐B/C, HLA‐A FHC products are also detected in individuals bearing HLA A3 and/or A30 allotypes. The aim of this study was to establish whether such restriction results in the detection of low FHC levels in individuals lacking HLA A3 and/or A30 allospecificities. The FHC mean concentration (± SD) in 294 healthy blood/bone marrow donors (HBDs) was 0.24 (± 0.2) mg/l. The grouping of HBDs according to their HLA‐A FHC product reactivity with one, both or no mAbs did not result in any statistically significant differences (Mann–Whitney test: P > 0.05) between their median FHC concentrations. Since the absence of differences in their FHC levels was not attributable to a difference in the percentage distribution of HLA allotypes associated with high or low HLA‐B/C FHC expression, our results indicate that FHC HLA A3 and/or A30 products detected in DDIA by these two mAbs only minimally contribute to FHC serum expression and that the assay is not limited by the failure to detect HLA‐A FHC products in A3− and/or A30− individuals.
Characterization of the NRAMP1 (SLC11A1) gene in the horse (Equus caballus L.)
Authors: Matiašovic J. , Kubíčková S. , Musilová P. , Rubeš J. , Hořín P.
Published: March 2003

The complete coding cDNA sequence of the horse NRAMP1 (SLC11A1) gene was determined (GenBank accession number AF354445). The nucleotide sequence of the horse NRAMP1 gene is similar to sequences of this gene in other species. The gene contains 15 exons whose total length of 1635 bp corresponds to 544 amino acids constituting the resulting putative protein. Hydrophobicity profile analysis of the deduced horse NRAMP1 gene product showed a nearly identical structure with the mouse NRAMP1 protein. The gene was found to be located on the short arm of ECA 6p12–13 by fluorescence in situ hybridization (FISH) analysis. Five allelic variants of the 5′ untranslated region (UTR) were identified at the nucleotide sequence level. PCR‐RFLP polymorphisms for NlaIII, TaqI, MspI and AciI were detected. Four out of five alleles could be detected using TaqI and MspI restriction enzymes. Their haplotype frequencies were different in four genetically distinct horse breeds.
Low frequency of HLA‐B*2706 in Taiwanese patients with ankylosing spondylitis
Authors: Chen I.‐H. , Yang K. L. , Lee A. , Huang H. H. , Lin P. Y. , Lee T. D.
Published: March 2003

The presence of HLA‐B27 in patients affected with ankylosing spondylitis (AS) was well established prior to the advent of DNA typing of various genes within the major histocompatibility complex (MHC) in humans. However, molecular typing of the MHC genes revealed that B27 comprises a motley assortment of alleles, some of which are strongly positively associated with the disease and some of which are negatively associated with the disease. B*2706 was reported to have a negative association with AS in the Thai population and in Chinese Singaporeans. We report here our finding of an absence of B*2706 in 184 Taiwanese AS patients.
Complete cDNA sequences of the HLA‐DRB1*0402 and DRB1*11041 alleles1
Authors: Ramon D. , Corell A. , Cox S. T. , Soteriou B. , Madrigal J. A. , Marsh S. G. E.
Published: March 2003

Since the development of the polymerase chain reaction, most HLA class II allele sequencing has been exclusively focused on the highly polymorphic exon 2. We present here the full cDNA sequences of two HLA‐DRB1 alleles, DRB1*0402 and DRB1*11041, both of which were previously only available as partial sequences. HLA‐DRB1*11041 was found to be completely homologous to DRB1*11011 in exons 1, 3, 4, 5 and 6 and HLA‐DRB1*0402 was found to be identical to DRB1*04011 in exons 1, 3, 4, 5 and 6.
Hotspot variation at the CH2–CH3 interface of leporid IgG antibodies (Oryctolagus, Sylvilagus and Lepus)1
Authors: Esteves P. J. , Alves P. C. , Ferrand N. , Van Der Loo W.
Published: March 2003

The European rabbit (Oryctolagus cuniculus) is the only species known to express only one subclass of gamma class immunoglobulin (IgG) antibodies. The rabbit IGHGCH2 (second domain of the gene region encoding the IgG heavy chain constant region) or e locus presents two serologically defined alleles, the e14 and e15 allotypes. These are correlated with amino acid variation at position 309, which is located within the target region of the neonatal FcRn receptor. The e14 and e15 markers were also observed in other lagomorph species. Population genetic research has indicated that polymorphism at this locus is sustained by selection. We present here the IGHGCH2 exon sequences for 12 species of rabbit and hare (genera Oryctolagus, Sylvilagus and Lepus). The inferred amino acid sequences reveal that, despite an overall sequence identity of 97%, five different residues can occur at position 309. As for Oryctolagus, the e15 allotype was always associated with the presence of an Ala309 codon. In all but one case, this codon defined an allotype‐specific ThaI restriction site. The potential of PCR/ThaI restriction fragment length polymorphism (RFLP) analyses for studying IGHGCH2 variation within and between populations is emphasized.
Forms of phosphorus transfer in hydrological pathways from soil under grazed grassland
Authors: Haygarth P. M. , Hepworth L. , Jarvis S. C.
Published: March 2003

Phosphorus (P) from soil can impair the water quality of streams and lakes. We have studied the forms and pathways of its movement from soil to water using 1‐ha plot lysimeters, managed as grazed grassland for 12 months in temperate South‐west England. The water flow through three pathways, namely (i) surface plus interflow to 30 cm (on undrained soil), (ii) surface plus interflow to 30 cm (on a mole and tile drained soil), and (iii) mole and tile drains (to 85 cm), were gauged. Samples of water from each path were treated with various combinations of 0.45‐μm filtration and sulphuric acid‐persulphate digestion and molybdate reaction, to determine the different forms of P. The total P (TP) concentration was greatest in the surface plus interflow to 30 cm paths (means 232 and 152 μg l–1), whereas the mean concentration in the drainage to 85 cm was 132 μg l–1. This reflects the substantial enrichment of the Olsen‐P extracts from the surface horizons, as extracts from the 0–2 cm layer were 10‐fold more than below 45 cm. In all paths, the dissolved P comprised the greatest proportion of the P transferred, with dissolved reactive P being the dominant form. Draining land reduced the transfer of TP by about 30% (≈ 1 kg–1 ha–1 year–1), because it can be sorbed as it flows through soil to drains. All these concentrations could cause eutrophication in surface waters.