bluuebunny/crossref_metadata_2025 huggingface dataset:
Differentiation‐dependent localisation of tissue‐type plasminogen activator in human bladder urothelium
Authors: Makuc Jana , Mašera Andrej , Tršinar Bojan , Jezernik Kristijan
Published: May 2004

Human bladder urothelium is able to secrete tissue‐type plasminogen activator (tPA). The aim of our study was to analyse localisation of tPA antigen in comparison to differentiation state of cells in samples of histologically normal urothelium and non‐invasive tumours of the human urinary bladder. Twenty‐five samples of normal urothelium and 31 non‐invasive papillary tumours from 36 patients were examined. The presence of tPA antigen was evaluated immunohistochemically. Differentiation of superficial cells was assessed by the presence of urothelial cell differentiation markers, uroplakins (UPs; immunohistochemistry) and cell's apical surface architecture (scanning electron microscopy). All tissue samples stained anti‐tPA positive. In normal urothelium, the intensity of anti‐tPA staining was the strongest in superficial cells, which were well‐differentiated. In tumours, all cell layers stained anti‐tPA positive. The intensity of anti‐tPA positive reaction in the upper cell layer correlated with the percentage of anti‐UP positive superficial cells. Superficial cells showed various differentiation states.

The localisation of tPA antigen in human in vivo tissue is not confined to the well‐differentiated superficial cells. Our results suggest a positive correlation between tPA secretion and cell differentiation.
A questionnaire‐based assessment of daily physical activity in heart failure
Authors: Garet Martin , Barthélémy Jean Claude , Degache Francis , Costes Frédéric , Da‐Costa Antoine , Isaaz Karl , Lacour Jean René , Roche Frédéric
Published: May 2004

Type and dose of daily energy expenditure (DEE) play a major role in modulations of health status and an increased knowledge of these dimensions of physical activity in congestive heart failure (CHF) patients would be valuable for clinical and epidemiological aims. We propose a new self‐administered DEE questionnaire adapted to CHF patients and tested its validity. One hundred and five stable CHF participants, NYHA class I–IV, LVEF=33.2±6.1% performed an incremental symptom‐limited Vo2 (peak) test and filled in the questionnaire for DEE calculation. Reproducibility (n=24), sensitivity (n=21) of the questionnaire and inter‐observer variability (n=105) were tested. Intensity levels were identified from DEE and their relationships to Vo2(peak), ventilatory and anthropometric characteristics were assessed by simple and multiple regression models. Reproducibility and sensitivity were high (r=0.98 and 0.88, respectively, P<0.0001) and inter‐observer error reached 1.37%. DEE was highly correlated to physical activity energy expenditure (r=0.96, P<0.0001). Relationships between DEE, Vo2(peak), VE/Vo2 and anthropometric characteristics were significant. An activity level above 3 MET was the best intensity criteria related to Vo2(peak) (r=0.62, P<0.0001) and DEE (r=0.80, P<0.0001). The questionnaire seems reproducible, sensible and valid for DEE estimation. Vo2(peak) appears related to DEE and especially to activities above 3 MET in CHF.
Selective intestinal decontamination in advanced chronic heart failure: a pilot trial
Authors: Conraads Viviane M. , Jorens Philippe G. , De Clerck Luc S. , Van Saene Hendrik K. , Ieven Margaretha M. , Bosmans Johan M. , Schuerwegh Annemie , Bridts Chris H. , Wuyts Floris , Stevens Wim J. , Anker Stefan D. , Rauchhaus Mathias , Vrints Christiaan J.
Published: June 2004

Background and aims:

Endotoxin, derived from intestinal aerobic Gram‐negative bacilli (AGNB), could be an important monocyte activator in chronic heart failure (CHF). The effect of selective decontamination of the digestive tract (SDD) on intracellular monocyte cytokine production, monocyte CD14 expression, circulating endotoxin and cytokines, and flow‐mediated dilation (FMD) was studied in patients with severe CHF.

Methods and results:

Ten patients with CHF (NYHA class III–IV) were enrolled in a non‐placebo controlled pilot trial involving the administration of SDD (polymyxin B, tobramycin) for 8 weeks. One patient was later excluded due to cardiac transplantation. Before treatment, after 4 and 8 weeks therapy, and 6 weeks post‐treatment, monocyte CD14 expression, intracellular monocyte production of interleukin‐1β [IL‐1β], interleukin‐6 [IL‐6], tumour necrosis factor (TNF)‐α with and without lipopolysaccharide (LPS) stimulation were measured. Concentrations of endotoxin and cytokines (IL‐1β, IL‐6, TNF‐α) were also determined. AGNB in faeces, intestinal endotoxin and FMD were assessed at baseline, after 4 weeks of treatment and 6 weeks post‐treatment. SDD eradicated intestinal AGNB (P<0.00001) and decreased faecal endotoxin concentrations (P<0.00001). There was a significant decline in monocyte CD14 expression (P=0.03) and in IL‐1β (P=0.0001), IL‐6 (P=0.02) and TNF‐α (P=0.0002) production after 4 and 8 weeks of treatment in the basal state and for IL‐1β (P=0.008) and IL‐6 (P=0.005) after LPS stimulation. FMD significantly improved at 4 weeks and returned to baseline after treatment discontinuation (P=0.002). Circulating concentrations of endotoxin and cytokines remained unchanged.

Conclusion:

Reduction of the intestinal endotoxin pool led to a decrease in monocyte CD14 expression and intracellular cytokine production in patients with severe CHF. The improvement of peripheral endothelial function could be a marker of the anti‐inflammatory effect of SDD.
P‐wave duration and P‐wave dispersion in patients with dilated cardiomyopathy
Authors: Senen Kubilay , Turhan Hasan , Erbay Ali Riza , Basar Nurcan , Yasar Ayse Saatci , Sahin Onur , Yetkin Ertan
Published: May 2004

Background:

P‐wave dispersion (PWD) has been reported to be associated with inhomogeneous and discontinuous propagation of sinus impulses. In the present study, we aimed to investigate PWD in patients with dilated cardiomyopathy.

Method:

The study population consisted of 72 patients with dilated cardiomyopathy and 72 healthy control subjects. Left atrial diameter, left ventricular end‐diastolic and end‐systolic diameters and left ventricular ejection fraction of all patients and control subjects were measured by means of transthoracic echocardiography. Maximum P‐wave duration (Pmaximum) and minimum P‐wave duration (Pminimum) were measured from the 12‐lead surface electrocardiogram. PWD was calculated as the difference between Pmaximum and Pminimum.

Results:

Pmaximum and PWD of patients with dilated cardiomyopathy were significantly higher than those of control subjects (Pmaximum: 126±12 ms vs. 116±10 ms, PWD: 47±6 ms vs. 38±7 ms, respectively, P<0.001 for all). However, there was no statistically significant difference between patient group and control group regarding Pminimum (79±7 ms vs. 78±6 ms, respectively, P=0.27). Left atrial diameter was significantly higher in patients with dilated cardiomyopathy compared to control subjects (4.51±0.62 cm vs. 3.60±0.43 cm, respectively, P<0.001). Left ventricular ejection fraction was found to be significantly lower in patients with dilated cardiomyopathy compared to control subjects (33±5% vs. 63±7%, respectively, P<0.001).

Conclusion:

PWD was found to be significantly higher in patients with dilated cardiomyopathy than in healthy control subjects.
Austrian survey of treating heart failure—AUSTRIA
Authors: Fruhwald Friedrich M. , Rehak Peter , Maier Robert , Watzinger Norbert , Wonisch Manfred , Klein Werner
Published: May 2004

Background:

Treatment of chronic heart failure is based on the results of large clinical trials, which form the basis of treatment guidelines, such as those from the European Society of Cardiology (ESC). The aim of this study was to record treatment‐modalities and the implementation of guidelines of chronic heart failure in clinical practice in Austria.

Methods:

Overall 96 general physicians, specialists for internal medicine in private practice or in hospital outpatient departments participated in the survey. Physicians were asked to prospectively document 30 consecutive patients with chronic heart failure.

Results:

1880 patients were documented. The majority of patients were treated by general physicians (57%). Coronary artery disease was the most frequent aetiology for heart failure (47%). The most frequently used drugs were blockers of the renin–angiotensin‐system (RAS‐blocker including ACE‐inhibitors and angiotensin‐receptor‐blockers, 78%), diuretics (76%) and beta‐blockers (49%). Other drugs like digitalis and spironolactone were used infrequently. Average doses of ACE‐inhibitors were approximately 90% of those recommended by the ESC, average doses of beta‐blockers were approximately 50% of those recommended. Treatment among the three classes of physicians differed with respect to RAS‐blockers and beta‐blockers, which were used infrequently by general practitioners. Both groups of drugs were given more frequently to younger patients (<70 years) while digitalis was given more often to elderly patients.

Conclusions:

Results from this survey suggest that Austrian physicians treating patients with heart failure use the appropriate drugs in dosages that are suggested by recently published guidelines (ACE‐inhibitors and beta‐blockers). However, dosages of spironolactone clearly differed from current recommendations.
Acute administration of diclofenac, but possibly not long term low dose aspirin, causes detrimental renal effects in heart failure patients treated with ACE‐inhibitors
Authors: Juhlin Tord , Björkman Sven , Gunnarsson Bodil , Fyge Åsa , Roth Bodil , Höglund Peter
Published: May 2004

Background:

Non‐steroidal anti‐inflammatory drugs (NSAID) or high doses of aspirin (acetylsalicylic acid) can exert detrimental effects on renal function and counteract the beneficial effects of angiotensin‐converting enzyme (ACE) inhibitors in patients with congestive heart failure.

Aims:

The objective of our study was to evaluate the renal effects of low dose aspirin and the NSAID diclofenac in patients with congestive heart failure treated with ACE‐inhibitors.

Methods:

Ten patients on their individually titrated dose of ACE‐inhibitors and low dose aspirin (≤125 mg daily) with stable congestive heart failure from coronary artery disease, entered an open investigation while on low dose aspirin, which was then discontinued. After one week wash‐out they received an oral dose of 50 mg diclofenac potassium or placebo in a double‐blind cross‐over fashion with a one week wash‐out period between treatments.

Results:

Diclofenac caused significant (P<0.05) decreases in GFR, urine flow, osmolality clearance, and excretion rates of sodium and potassium compared to placebo and aspirin. At tmax for diclofenac or corresponding time for placebo diclofenac caused 40 (11–59)% (geometric mean and 95% confidence limits) reduction in GFR compared to placebo and 36 (5.4–56)% reduction to low‐dose aspirin. No significant changes between low dose aspirin and placebo were found.

Conclusion:

Acute administration of diclofenac, but not long term low dose aspirin, has profound impact on renal function in patients with heart failure treated with ACE‐inhibitors and may cause worsened heart failure.
An unusual cause of heart failure identified by echocardiography in an octogenarian
Authors: Lin Ming Lee , Connelly Kim , Prior David
Published: May 2004

Non‐compaction of the ventricular myocardium is a rare congenital cardiomyopathy that carries an unfavorable prognosis. It is associated with a high incidence of progressive heart failure, thromboembolism and malignant arrhythmias. Echocardiography currently remains the imaging modality most commonly used for diagnosis. We describe an unusual case of isolated non‐compaction of the left ventricle in an octogenarian male to highlight the need for echocardiography in all patients with suspected heart failure, in order to determine aetiology, prognosis and assess treatment.
Short‐term prognostic value of initial serum levels of interleukin‐10 in patients with acute myocarditis
Authors: Fuse Koichi , Kodama Makoto , Okura Yuji , Ito Masahiro , Kato Kiminori , Hanawa Haruo , Aizawa Yoshifusa
Published: May 2004

The disease course of acute myocarditis has a wide spectrum and the predictors of the prognosis in patients with acute myocarditis have not yet been established. In the pathogenesis of myocarditis, the cytokine environment is important. In this study, we examined the predictive values of serum levels of interleukin‐10 (IL‐10) and IL‐12 in the short‐term prognosis of patients with acute myocarditis. Twenty‐four consecutive patients who had been diagnosed as having acute active myocarditis were analyzed and monitored for 2 months. The patients with myocarditis were divided into the survival group (n=16) and the non‐survival group (n=8). Initial serum levels of IL‐10 (P=0.0015) and IL‐12 (P=0.012) in the non‐survival group were significantly higher than those of the survival group, and there was a significant correlation between IL‐10 and IL‐12 levels (P<0.0001). The univariate analyses showed that increased serum levels of IL‐10 (hazard ratio 1.041, P=0.0004) and IL‐12 (hazard ratio 1.128, P=0.0346) were significant predictors of mortality. In the Kaplan–Meier analysis, high levels of IL‐10 (≥7.0 pg/ml) (P=0.0239) strongly predicted high mortality. In conclusion, the elevation in serum IL‐10 levels at the initial phase appeared to predict poor short‐term prognosis in patients with acute myocarditis.
Hereditary hemochromatosis gene (HFE) mutations C282Y, H63D and S65C in patients with idiopathic dilated cardiomyopathy
Authors: Hannuksela Jokke , Leppilampi Mari , Peuhkurinen Keijo , Kärkkäinen Satu , Saastamoinen Eija , Heliö Tiina , Kaartinen Maija , Nieminen Markku S. , Nieminen Pentti , Parkkila Seppo
Published: May 2004

Background:

Hereditary hemochromatosis (HH), a common autosomal recessive disease, leads to excessive iron accumulation in some organs, including the heart. It is therefore not surprising that cardiomyopathy is one of the most severe complications of HH. The HFE gene defects have been thought to contribute to idiopathic dilated cardiomyopathy (IDCM) in some patients, even though the results of genotype analyses have so far been contradictory. Hence we set out here to evaluate the prevalence and potential role of HFE mutations in patients with IDCM.

Methods:

A total of 91 IDCM patients and 102 controls were subjected to HFE mutation analyses, in which C282Y, H63D and S65C mutations were determined for each patient. We also analyzed the impact of the C282Y and H63D mutations on the left ventricular end‐diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF) and New York Heart Association (NYHA) functional classes.

Results:

The prevalences of heterozygosity for the C282Y, H63D and S65C mutations in the IDCM patients were 13.2%, 22.0% and 2.2%, respectively. LVEDD was significantly higher (P=0.037) in those with the C282Y mutation at the end of the follow‐up period than in those with no mutation.

Conclusions:

Our data showed no significant deviations in C282Y, H63D and S65C mutation frequencies between the IDCM patients and controls, suggesting that these mutations do not increase the risk of IDCM. Heterozygosity for the C282Y mutation may nevertheless be a modifying factor contributing to LV dilatation and remodeling.
Natriuretic peptides in the monitoring of anthracycline induced reduction in left ventricular ejection fraction
Authors: Daugaard Gedske , Lassen Ulrik , Bie Peter , Pedersen Erling Bjerregaard , Jensen Kaare Troels , Abildgaard Ulrik , Hesse Birger , Kjaer Andreas
Published: May 2004

Background:

The use of anthracyclines in treatment of cancer is limited by cardiotoxicity of these compounds and may lead to heart failure. Therefore monitoring of cardiac function is necessary during therapy.

Aim:

We evaluated the value of natriuretic peptides (N‐terminal pro‐atrial natriuretic peptide (N‐ANP) and brain natriuretic peptide (BNP)) for monitoring and predicting anthracycline induced cardiotoxicity using radionuclide left ventricular ejection fraction (EF) measurements as reference.

Methods and results:

A total of 107 consecutive patients receiving anthracycline as part of their chemotherapy for malignant disease were studied. Plasma concentrations of the peptides were measured by radioimmunoassay and EF by radionuclide cardiography. For reduced EF values, i.e. below 0.50 a fairly strong correlation was found between N‐ANP or BNP and EF. Of 48 patients with serial EF and peptide measurements, 19% showed a significant EF decrease (>0.10) and ended with a final EF value below 0.50. Baseline EF was no predictor of a change in EF during treatment. Neither baseline levels of N‐ANP or BNP nor a change in the same variables during therapy were predictive of a change in EF.

Conclusions:

In spite of correlations between peptide concentrations and reduced EF values neither baseline values nor serial measurements can safely substitute EF monitoring in patients undergoing anthracycline therapy.
Clinical trials update and cumulative meta‐analyses from the American College of Cardiology: WATCH, SCD‐HeFT, DINAMIT, CASINO, INSPIRE, STRATUS‐US, RIO‐Lipids and cardiac resynchronisation therapy in heart failure
Authors: Cleland John G.F. , Ghosh Justin , Freemantle Nick , Kaye Gerry C. , Nasir Mansoor , Clark Andrew L. , Coletta Alison P.
Published: June 2004

This article continues a series of reports on recent research developments in the field of heart failure. Key presentations made at the American College of Cardiology meeting, held in New Orleans, Louisiana, USA in March 2004 are reported. These new data have been added to existing data in cumulative meta‐analyses. The WATCH study randomised 1587 patients with heart failure and left ventricular systolic dysfunction to warfarin, aspirin or clopidogrel. The study showed no difference between the effects of these agents on mortality or myocardial infarction, but hospitalisations for heart failure were higher on aspirin (22.2%) compared to warfarin (16.1%). The SCD‐HeFT study showed that ICD therapy reduced all‐cause mortality at 5 years by 23% in patients with predominantly NYHA class II heart failure and left ventricular systolic dysfunction, but amiodarone was ineffective. The DINAMIT study showed that ICD therapy was not beneficial in patients with left ventricular dysfunction after a recent MI, even in those with risk factors for arrhythmic death. In CASINO, levosimendan improved survival compared with dobutamine or placebo in patients with decompensated heart failure. INSPIRE showed that SPECT imaging can be used to assess risk early after acute MI safely and accurately. Rimonabant was shown to be safe and effective in treating the combined cardiovascular risk factors of smoking and obesity. An overview of new developments in cardiac resynchronisation therapy (CRT) in heart failure is also reported.
Circadian clocks and natural antisense RNA
Authors: Crosthwaite Susan K.
Published: May 2004

Eukaryotes regulate gene expression in a number of different ways. On a daily and seasonal timescale, the orchestration of gene expression is to a large extent governed by circadian clocks. These endogenous timekeepers enable organisms to prepare for predictable environmental conditions from one day to the next and thus allow adaptation to a given temporal niche. In general, circadian clocks have been shown to employ the classical transcriptional and posttranscriptional control mechanisms to generate rhythmicity. However, the discovery of antisense clock gene transcripts suggests that mechanisms of gene regulation operating through antisense RNA may also be integral to the circadian clockwork. Following a brief history of the impact of genetic and molecular techniques in aiding our understanding of circadian clocks, this review concentrates on the few examples of antisense clock gene transcripts so far investigated and their effect on circadian timing.
Endoreduplication and activation of the anaphase‐promoting complex during symbiotic cell development
Authors: Kondorosi Eva , Kondorosi Adam
Published: May 2004

Postembryonic development of plant organs requires a constant interplay between the cell cycle and the developmental programs. Upon endo‐ and exogenous signals, plant cells can enter, exit or modify the cell cycle. Alteration of mitotic cycles to endoreduplication cycles, where the genome is duplicated without mitosis, is common in plants and may play a role in cell differentiation. The switch from the mitotic to endocycles is regulated by Ccs52A, a plant orthologue of the yeast and animal Cdhl proteins, acting as substrate‐specific activator of the anaphase‐promoting complex E3 ubiquitin ligase. Here, several aspects of endoreduplication are discussed with special attention on nitrogen‐fixing nodule development where endoreduplication is an integral part of symbiotic cell differentiation.
Happy birthday: forty years of FEBS
Authors: Feldmann Horst
Published: May 2004

The 2004 Federation of European Biochemical Societies (FEBS) Congress in Warsaw marks the 40th Anniversary of FEBS. To celebrate this event, the Executive Committee decided to publish a memoir, which chronicles the foundation of FEBS and its early development as well as presents an overview of FEBS activities and contributions that support the ever growing disciplines of biochemistry, molecular cell biology and molecular biophysics throughout Europe [Forty Years of FEBS Horst Feldmann (Ed.), Blackwell Publishers, Oxford 2003, on behalf of FEBS]. This paper summarizes some of the most important aspects of this compilation.
Regulatory mechanisms controlling biogenesis of ubiquitin and the proteasome
Authors: London Markus K. , Keck Birgit I. , Ramos Paula C. , Jürgen Dohmen R.
Published: June 2004

Analysis of several Saccharomyces cerevisiae ump mutants with defects in ubiquitin (Ub)‐mediated proteolysis yielded insights into the regulation of the polyubiquitin gene UBI4 and of proteasome genes. High‐molecular weight Ub–protein conjugates accumulated in ump mutants with impaired proteasome function with a concomitant decrease in the amount of free Ub. In these mutants, transcriptional induction of UBI4 was depending in part on the transcription factor Rpn4. Deletion of UBI4 partially suppressed the growth defects of ump1 mutants, indicating that accumulation of polyubiquitylated proteins is deleterious to cell growth. Transcription of proteasome subunit genes was induced in ump mutants affecting the proteasome, as well as under conditions that mediate DNA damage or the formation of abnormal proteins. This induction required the transcriptional activator Rpn4. Elevated Rpn4 levels in proteasome‐deficient mutants or as a response to abnormal proteins were due to increased metabolic stability. Up‐regulation of proteasome genes in response to DNA damage, in contrast, is shown to operate via induction of RPN4 transcription.
Overexpression of wild‐type and mutant mucolipin proteins in mammalian cells: effects on the late endocytic compartment organization
Authors: Manzoni M. , Monti E. , Bresciani R. , Bozzato A. , Barlati S. , Bassi M.T. , Borsani G.
Published: June 2004

Mucolipin‐1 is a 65‐kDa membrane protein encoded by the MCOLN1 gene, which is mutated in patients with mucolipidosis type IV (MLIV), a rare neurodegenerative lysosomal storage disorder. We studied the subcellular localization of wild‐type and three different mutant forms (T232P, F408del and F465L) of mucolipin by expressing Myc‐tagged proteins in HeLa cells. The overexpressed wild‐type mucolipin colocalizes to late endocytic structures and induces an aberrant distribution of these compartments. F408del and F465L MLIV mutant proteins show a distribution similar to the wild‐type protein, whereas T232P is retained in the endoplasmic reticulum. Among the mutants, only F408del induces a redistribution of the late endocytic compartment. These findings suggest that the overexpression of the mucolipin cation channel influences the dynamic equilibrium of late endocytic compartments.
Poorer NF‐κB signaling by microfilariae in macrophages from BALB/c mice affects their ability to produce cytotoxic levels of nitric oxide to kill microfilariae
Authors: Mukhopadhyay Sangita , Srivastava V.M.L. , Murthy P.K. , Hasnain Seyed E.
Published: June 2004

Upon activation with microfilariae (mf), macrophages from C57Bl/6 mice showed higher nuclear factor‐κB (NF‐κB) but lower activating protein 1 DNA‐binding activity as compared to BALB/c macrophages. The C57Bl/6 macrophages produced cytotoxic levels of nitric oxide (NO) to kill Setaria cervi mf as compared to BALB/c macrophages. Inhibition of the NF‐κB signal by pyrrolidine dithiocarbamate (PDTC) blocked NO production and microfilaricidal activity of C57Bl/6 macrophages and inclusion of the exogenous NO generator (SNP) in the PDTC treated C57Bl/6 macrophage cultures induced mf cytotoxicity. These results underscore that the NF‐κB signal (induced in response to mf) is important for the NO‐mediated microfilaricidal activity of macrophages.
Study of cytokine induced neuropathology by high resolution proton NMR spectroscopy of rat urine
Authors: Griffin Julian L , Anthony Daniel C , Campbell Sandra J , Gauldie Jack , Pitossi Fernando , Styles Peter , Sibson Nicola R
Published: May 2004

Multiple sclerosis is a major cause of non‐traumatic neurological disability. The identification of markers that differentiate disease progression is critical to effective therapy. A combination of NMR spectroscopic metabolic profiling of urine and statistical pattern recognition was used to detect focal inflammatory central nervous system (CNS) lesions induced by microinjection of a replication‐deficient recombinant adenovirus expressing TNF‐α or IL1‐β cDNA into the brains of Wistar rats. These animals were compared with a group of naı̈ve rats and a group of animals injected with an equivalent null adenovirus. Urine samples were collected 7 days after adenovirus injection, when the inflammatory lesion is maximally active. Principal components analysis and Partial Least Squares‐Discriminate analysis of the urine 1H NMR spectra revealed significant differences between each of the cytokine adenovirus groups and the control groups; for the TNF‐α group the main differences lay in citrate and succinate, while for the IL‐1β group the predominant changes occurred in leucine, isoleucine, valine and myo‐inositol. Thus, we can identify urinary metabolic vectors that not only separate rats with inflammatory lesions in the brain from control animals, but also distinguish between different types of CNS inflammatory lesions.
Detection of UDP–glucose:cyclo‐DOPA 5‐O‐glucosyltransferase activity in four o'clocks (Mirabilis jalapa L.)
Authors: Sasaki Nobuhiro , Adachi Taiji , Koda Takatoshi , Ozeki Yoshihiro
Published: May 2004

Although a pathway for betacyanin biosynthesis has been postulated, most of the catalytic steps have not yet been identified or demonstrated with biochemical evidence. In the postulated pathway, the glucose moiety of betanin is conjugated to the aglycone, betanidin, because the glucosyltransferase (GT) activity that produces betanin has been reported and its cDNA isolated. However, another pathway for betacyanin biosynthesis is proposed in which betanin is formed by GT acting at the 5,6‐dihydroxyindoline‐2‐carboxylic acid (cyclo‐DOPA) step, followed by condensation of the product with betalamic acid. Here, we show that GT activity acts upon cyclo‐DOPA in the betacyanin synthetic pathway. A crude extract from the petals of four o'clocks (Mirabilis jalapa L.) was mixed with cyclo‐DOPA and UDP–glucose. After the reaction was stopped with phosphoric acid, the product was chemically reacted with betalamic acid. In the final reaction mixture, betanin formation was confirmed by HPLC analysis, demonstrating cyclo‐DOPA 5‐O‐glucosyltransferase activity. This activity was correlated with the accumulation of betanin during the development of four o'clock flowers and was detected in another five species of Centrospermae. These results indicate that the glucose moiety of betanin is introduced at the cyclo‐DOPA step, which is followed by condensation with betalamic acid, and not at the betanidin aglycone step.
Amino acid type selective isotope labelling of the multidrug ABC transporter LmrA for solid‐state NMR studies
Authors: James Mason A , Siarheyeva Alena , Haase Winfried , Lorch Mark , van Veen Hendrik , Glaubitz Clemens
Published: May 2004

The ABC transporter LmrA in Lactococcus lactis confers resistance to a wide range of antibiotics and cytotoxic drugs and is a functional homologue of P‐glycoprotein. Recently, solid‐state NMR methods have shown potential for structural‐ and non‐perturbing, site directed functional studies. These experiments require isotopic labelling of selected sites. We have developed a strategy to produce large quantities of selectively labelled LmrA reconstituted at a high density in lipid membranes. This makes the 64 kDa integral membrane protein LmrA and therefore the ABC transporter superfamily accessible to NMR analysis.
Two different dihydroorotate dehydrogenases from yeast Saccharomyces kluyveri
Authors: Zameitat Elke , Knecht Wolfgang , Piškur Jure , Löffler Monika
Published: May 2004

Genes for two structurally and functionally different dihydroorotate dehydrogenases (DHODHs, EC 1.3.99.11), catalyzing the fourth step of pyrimidine biosynthesis, have been previously found in yeast Saccharomyces kluyveri. One is closely related to the Schizosaccharomyces pombe mitochondrial family 2 enzymes, which use quinones as direct and oxygen as the final electron acceptor. The other one resembles the Saccharomyces cerevisiae cytosolic family 1A fumarate‐utilizing DHODH. The DHODHs from S. kluyveri, Sch. pombe and S. cerevisiae, were expressed in Escherichia coli and compared for their biochemical properties and interaction with inhibitors. Benzoates as pyrimidine ring analogs were shown to be selective inhibitors of cytosolic DHODs. This unique property of Saccharomyces DHODHs could appoint DHODH as a species‐specific target for novel anti‐fungal therapeutics.
Microautophagy and macropexophagy may occur simultaneously in Hansenula polymorpha
Authors: Monastryska Iryna , Sjollema Klaas , van der Klei Ida J , Kiel Jan A.K.W , Veenhuis Marten
Published: May 2004

We subjected methanol‐grown cells of wild type Hansenula polymorpha simultaneously to nitrogen depletion and excess glucose conditions. Both treatments induce the degradation of peroxisomes, either selective (via excess glucose) or non‐selective (via nitrogen limitation). Our combined data strongly suggest that both processes occur simultaneously under these conditions. The implications of these findings on studies of autophagy and related transport pathways to the vacuole in yeast are discussed.
Cell‐type‐specific interactions at regulatory motifs in the first intron of the lamin A gene
Authors: Arora Puneeta , Muralikrishna Bh , Parnaik Veena K
Published: May 2004

Lamins A, C and C2 are alternatively spliced products of the LMNA gene; lamins A and C are expressed in differentiated somatic cells, whereas lamin C2 is expressed in germ cells. We have analyzed a segment of the first intron of the LMNA gene for cell‐type‐specific regulatory elements. We identified a 420‐bp fragment that increased promoter activity in lamin A‐expressing cells but repressed activity in undifferentiated cells. DNase I footprinting and electrophoretic mobility shift assays revealed two binding motifs, footprinted region A (FPRA) and FPRB. The hepatocyte nuclear factor‐3β was bound to FPRA only in somatic cell extracts and this motif had an inhibitory effect on promoter activity. The retinoic X receptor β, RXRβ, bound near FPRB with extracts from lamin A‐ or C2‐expressing cells, and this site enhanced promoter activity. We have, thus, identified two novel binding sites for transcription factors in a region likely to function as an important regulatory element for the cell‐type‐specific transcription of A‐type lamins.
Overexpression of GD3 synthase induces apoptosis of vascular endothelial ECV304 cells through downregulation of Bcl‐2
Authors: Ha Ki-Tae , Lee Young-Choon , Kim Cheorl-Ho
Published: May 2004

The disialoganglioside GD3 plays a major role in proliferation, differentiation, and apoptosis. It has been reported that ganglioside GD3 can induce apoptosis through bcl‐2 mediated mitochondrial pathway. However, the relationship between ganglioside GD3 and B‐cell/CLL lymphoma 2 (Bcl‐2) is not fully understood. In this study, we have demonstrated that the downregulation of Bcl‐2 by overexpression of CMP‐NeuAc:GM3 α‐2,8‐sialyltransferase (GD3 synthase) results in an accelerated apoptosis in vascular endothelial cells (ECV304), as evidenced by DNA fragmentation and caspase‐3 activation. In addition, phosphorylation of AKT and cyclic‐AMP responsive element binding protein (CREB) was reduced by GD3 synthase overexpression. Moreover, the activation of CREB as a transcriptional factor was also inhibited, as evidenced by electrophoretic mobility shift assay. Therefore, we conclude that GD3 synthase has an apoptotic effect on ECV304 cells through downregulation of Bcl‐2 expression via dephosphorylation of AKT and CREB.
Aquaporin‐2 trafficking is regulated by PDZ‐domain containing protein SPA‐1
Authors: Noda Yumi , Horikawa Saburo , Furukawa Tetsushi , Hirai Keiji , Katayama Yoshifumi , Asai Tomoki , Kuwahara Michio , Katagiri Koko , Kinashi Tatsuo , Hattori Masakazu , Minato Nagahiro , Sasaki Sei
Published: May 2004

Targeted positioning of water channel aquaporin‐2 (AQP2) strictly regulates body water homeostasis. Trafficking of AQP2 to the apical membrane is critical to the reabsorption of water in renal collecting ducts. Controlled apical positioning of AQP2 suggests the existence of proteins that interact with AQP2. A biochemical search for AQP2‐interacting proteins led to the identification of PDZ‐domain containing protein, signal‐induced proliferation‐associated gene‐1 (SPA‐1) which is a GTPase‐activating protein (GAP) for Rap1. The distribution of SPA‐1 coincided with that of AQP2 in renal collecting ducts. The site of colocalization was concomitantly relocated by hydration status. AQP2 trafficking to the apical membrane was inhibited by the SPA‐1 mutant lacking Rap1GAP activity and by the constitutively active mutant of Rap1. AQP2 trafficking was impaired in SPA‐1‐deficient mice. Our results show that SPA‐1 directly binds to AQP2 and regulates at least in part AQP2 trafficking.
A mechanism of impaired mobility of oligodendrocyte progenitor cells by tenascin C through modification of wnt signaling
Authors: Kakinuma Yoshihiko , Saito Fumiji , Osawa Shizue , Miura Masayuki
Published: May 2004

In demyelinating diseases, the mechanisms of how oligodendrocyte (OLG) progenitor cells are affected in the demyelinated area remain to be elucidated. To investigate one aspect of the mechanisms, we focused on the role of tenascin C in regulating the migratory mobility of the progenitor cells via β‐catenin. By cDNA subtraction screening, we found tenascin C expression to be increased in OLG progenitors (rat primary O2A cells). Tenascin C inhibited the migration of OLG progenitors and CG‐4 cells, and β‐catenin accumulated at focal adhesions in these cells. These changes were associated with the inactivation of canonical wnt signaling. Overexpression of the wnt‐signaling antagonist Dapper prevented the migration of CG‐4 cells. This suggests that inactivation of the wnt signal is responsible for impaired migration of OLG caused by tenascin C. Our results suggest that tenascin C is involved in the impaired mobility of OLG progenitor cells through increased amounts of adhesion complex as well as the prevention of wnt signaling.
Voltage sensor mutations differentially target misfolded K+ channel subunits to proteasomal and non‐proteasomal disposal pathways
Authors: Myers Michael P , Khanna Rajesh , Lee Eun Jeon , Papazian Diane M
Published: May 2004

In Shaker K+ channels, formation of an electrostatic interaction between two charged residues, D316 and K374 in transmembrane segments S3 and S4, respectively, is a key step in voltage sensor biogenesis. Mutations D316K and K374E disrupt formation of the voltage sensor and lead to endoplasmic reticulum retention. We have now investigated the fates of these misfolded proteins. Both are significantly less stable than the wild‐type protein. D316K is degraded by cytoplasmic proteasomes, whereas K374E is degraded by a lactacystin‐insensitive, non‐proteasomal pathway. Our results suggest that the D316K and K374E proteins are misfolded in recognizably different ways, an observation with implications for voltage sensor biogenesis.
Dihydrolipoamide dehydrogenase from porcine heart catalyzes NADH‐dependent scavenging of nitric oxide
Authors: Igamberdiev Abir U , Bykova Natalia V , Ens Werner , Hill Robert D
Published: May 2004

Dihydrolipoamide dehydrogenase (DLDH; EC 1.8.1.4) from porcine heart is capable of using nitric oxide (NO) as an electron acceptor, with NADH as the electron donor, forming nitrate in the reaction. NADPH was not effective as an electron donor. The reaction had a pH optimum near 6 and was not inhibited by cyanide or diphenyleneiodonium ions. The K
m for NADH was 10 μM, while that for NO was 0.5 μM. The rate of NO conversion was comparable to the rate of lipoamide conversion (200 μmol min−1 mg−1 protein at pH 6). Cytochrome c or myoglobin were poor electron acceptors by themselves but, in the presence of methylene blue, DLDH had an activity of 5–7 μmol min−1 mg−1 protein with these substrates, indicating that DLDH can act also as a methemoglobin reductase. While the K
m of DLDH for NO is relatively low, it is in the physiological range of NO levels encountered in the tissue. The enzyme may, therefore, have a significant role in modifying NO levels under specific cell conditions.
Functional chimeras between the catalytic domains of the mycobacterial adenylyl cyclase Rv1625c and a Paramecium guanylyl cyclase
Authors: Linder Jürgen U , Castro Lucila I , Guo Ying-Lan , Schultz Joachim E
Published: May 2004

The class IIIa adenylyl cyclase (AC) Rv1625c from Mycobacterium tuberculosis forms homodimers with two catalytic centres, whereas the Paramecium guanylyl and mammalian ACs operate as pseudoheterodimers with one catalytic centre. The functional and structural relationship of the catalytic domains of these related class III cyclases was investigated. Point mutations introduced into Rv1625c to engineer a forskolin‐binding pocket created a single heterodimeric catalytic centre, yet did not result in forskolin activation. Chimerization of these Rv1625c point mutants with corresponding mammalian AC domains was impossible. However, it was successful using a complemental Paramecium guanylyl cyclase domain and resulted in an AC. The data signify a divergence of structural and functional evolution in class III Acs.
The mutual information theory for the certification of rice coding sequences
Authors: Carels Nicolas , Vidal Ramon , Mansilla Ricardo , Frı́as Diego
Published: May 2004

We report here the use of the mutual information theory for the certification of annotated rice coding sequences of both GenBank and TIGR databases. Considering coding sequences larger than 600 bp, we successfully screened out genes with aberrant compositional features. We found that they represent about 10% of both datasets after cleaning for gene redundancy. Most of the rejected accessions showed a different trend in GC3% vs GC2% plot compared to the set of accessions that have been published in international journals. This suggests the existence of a bias in the pattern recognition algorithms used by gene prediction programs.
Receptor binding specificities of Herstatin and its intron 8‐encoded domain
Authors: Shamieh Lara S , Evans Adam J , Denton Michael C , Clinton Gail M
Published: May 2004

Retention of intron 8 in alternative HER‐2 mRNA generates an inhibitory secreted ligand, Herstatin, with a novel receptor‐binding domain (RBD) encoded by the intron. This study examines binding interactions with several receptors and investigates sequence variations in intron 8. The RBD, expressed as a peptide, binds at nM concentrations to HER‐2, the EGFR, ΔEGFR, HER‐4 and to the IGF‐1 receptor, but not to HER‐3 nor to the FGF‐3 receptor, whereas a rare mutation in the RBD (Arg to Ile) eliminates receptor binding. The full‐length Herstatin binds with 3–4‐fold higher affinity than its RBD, but with ∼10‐fold lower affinity to the IGF‐IR. Sequence conservation in rhesus monkey but not in rat suggests that intron 8 recently diverged as a receptor‐binding module critical for the function of Herstatin.
Manual removal or spontaneous placental delivery and postcesarean endometritis and bleeding
Authors: Dehbashi S , Honarvar M , Fardi F.H
Published: May 2004

Objectives: To compare the effect of manual removal and spontaneous delivery of placenta on postcesarean bleeding and endometritis. Methods: In this prospective study 400 pregnant women undergoing elective cesarean delivery were randomly assigned to two groups: spontaneous placental delivery (200 women) and manual placental delivery (200). No antibiotic prophylaxis was administered in either group. The significance of blood loss due to cesarean delivery was defined by a drop in hemoglobin concentration of more than 1 g/dl 24 h after delivery in comparison with preoperative hemoglobin concentration. Endometritis was diagnosed in patients who developed clinical signs of fever and suprapubic tenderness 48 h after delivery. We analyzed the data using a t‐test. Results: The overall endometritis rate was 27%, in 40 women in the spontaneous placental delivery group (20%) and 68 women in the manual placental removal group (34%). There was a statistically significant difference between the two groups (P=0.001). Significant blood loss was experienced by 52 women (26%) in the spontaneous placental delivery group vs. 100 women (50%) in the manual placental delivery group. There was a statistically significant difference in blood loss between the two groups (P=0.000; RR=1.92). Conclusion: Endometritis and blood loss in cesarean delivery is increased by manual removal of the placenta compared with the spontaneous method of placental removal.
Chronic hypertension and the risk for adverse pregnancy outcome after superimposed pre‐eclampsia
Authors: Vanek M , Sheiner E , Levy A , Mazor M
Published: May 2004

Objective: To determine the risk factors and pregnancy outcome of patients with chronic hypertension during pregnancy after controlling for superimposed preeclampsia. Method: A comparison of all singleton term (>36 weeks) deliveries occurring between 1988 and 1999, with and without chronic hypertension, was performed. Stratified analyses, using the Mantel–Haenszel technique, and a multiple logistic regression model were performed to control for confounders. Results: Chronic hypertension complicated 1.6% (n=1807) of all deliveries included in the study (n=113 156). Using a multivariable analysis, the following factors were found to be independently associated with chronic hypertension: maternal age >40 years (OR=3.1; 95% CI 2.7–3.6), diabetes mellitus (OR=3.6; 95% CI 3.3–4.1), recurrent abortions (OR=1.5; 95% CI 1.3–1.8), infertility treatment (OR=2.9; 95% CI 2.3–3.7), and previous cesarean delivery (CD; OR=1.8 CI 1.6–2.0). After adjustment for superimposed preeclampsia, using the Mantel–Haenszel technique, pregnancies complicated with chronic hypertension had higher rates of CD (OR=2.7; 95% CI 2.4–3.0), intra uterine growth restriction (OR=1.7; 95% CI 1.3–2.2), perinatal mortality (OR=1.6; 95% CI 1.01–2.6) and post‐partum hemorrhage (OR=2.2; 95% CI 1.4–3.7). Conclusion: Chronic hypertension is associated with adverse pregnancy outcome, regardless of superimposed preeclampsia.
Beyond Belief: Religious Experience, Ritual, and Cultural Neuro-phenomenology in the Interpretation of Past Religious Systems
Authors: Dornan Jennifer L.
Published: June 2004

While there is growing agreement within anthropology and archaeology that notions of ‘experience’ can contribute to our interpretations of the past, this article suggests that there is a need to incorporate insight gathered from the fields of cultural phenomenology and cultural neuro-phenomenology into general anthropological understandings of cross-cultural religious experience. Specifically, this article explores the insight offered by cultural neuro-phenomenology into the relationships between religious symbolism, ritual, power, religious belief, and individual religious experience. In assessing the role that belief, as instantiated through ritually-induced religious experience, plays in the maintenance or alteration of state-level religious systems, this article will outline the ways in which this insight may both help us better to understand past religious experience as well as to interpret the maintenance and alteration of past religious systems. To demonstrate the potential of this approach, this article will conclude with a brief discussion of the fall of the Classic Maya state religious system.
A Cognitive Typology for Numerical Notation
Authors: Chrisomalis Stephen
Published: June 2004

The numerical notation associated with texts and other representational media used in ancient societies is an important means by which past cognitive processes may be reconstructed. No satisfactory typology exists, however, to help understand the relationship between numerical symbols and cognitive processes. As a result, theories concerning the development of numeration remain mired in a unilinear and ethnocentric framework in which our own (Hindu-Arabic or Western) numerals are seen as the ultimate stage of evolution. It is suggested herein that there are two separate dimensions that need to be considered when classifying and evaluating numerical notation systems, and that these dimensions are structured in highly constrained ways. A new typology is presented in which systems are classified into five major types on the basis of these dimensions. Using this typology, a multilinear model is presented for the patterned diachronic change in numerical notation systems, which refutes both unilinear evolutionary theories and radically relativistic propositions regarding how individuals in pre-modern societies represented numbers.
Palaeolithic Populations and Waves of Advance
Authors: Fort Joaquim , Pujol Toni , Cavalli-Sforza Luigi Luca
Published: June 2004

The wave-of-advance model has been previously applied to Neolithic human range expansions, yielding good agreement to the speeds inferred from archaeological data. Here, we apply it for the first time to Palaeolithic human expansions by using reproduction and mobility parameters appropriate to hunter-gatherers (instead of the corresponding values for preindustrial farmers). The order of magnitude of the predicted speed is in agreement with that implied by the AMS radiocarbon dating of the lateglacial human recolonization of northern Europe (14.2–12.5 kyr bp). We argue that this makes it implausible for climate change to have limited the speed of the recolonization front. It is pointed out that a similar value for the speed can be tentatively inferred from the archaeological data on the expansion of modern humans into the Levant and Europe (42–36 kyr bp).