bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular and Genetic Determinants of Rous Sarcoma Virus Integrase for Concerted DNA Integration
Authors: Chiu Roger , Grandgenett Duane P.
Published: May 2003

Site-directed mutagenesis of recombinant Rous sarcoma virus (RSV) integrase (IN) allowed us to gain insights into the protein-protein and protein-DNA interactions involved in reconstituted IN-viral DNA complexes capable of efficient concerted DNA integration (termed full-site). At 4 nM IN, wild-type (wt) RSV IN incorporates ∼30% of the input donor into full-site integration products after 10 min of incubation at 37°C, which is equivalent to isolated retrovirus preintegration complexes for full-site integration activity. DNase I protection analysis demonstrated that wt IN was able to protect the viral DNA ends, mapping ∼20 bp from the end. We had previously mapped the replication capabilities of several RSV IN mutants (A48P and P115S) which appeared to affect viral DNA integration in vivo. Surprisingly, recombinant RSV A48P IN retained wt IN properties even though the virus carrying this mutation had significantly reduced integrated viral DNA in comparison to wt viral DNA in virus-infected cells. Recombinant RSV P115S IN also displayed all of the properties of wt RSV IN. Upon heating of dimeric P115S IN in solution at 57°C, it became apparent that the mutation in the catalytic core of RSV IN exhibited the same thermolabile properties for 3′ OH processing and strand transfer (half-site and full-site integration) activities consistent with the observed temperature-sensitive defect for integration in vivo. The average half-life for inactivation of the three activities were similar, ranging from 1.6 to 1.9 min independent of the IN concentrations in the assay mixtures. Wt IN was stable under the same heat treatment. DNase I protection analysis of several conservative and nonconservative substitutions at W233 (a highly conserved residue of the retrovirus C-terminal domain) suggests that this region is involved in protein-DNA interactions at the viral DNA attachment site. Our data suggest that the use of recombinant RSV IN to investigate efficient full-site integration in vitro with reference to integration in vivo is promising.
Cellular Gene Expression Survey of Vaccinia Virus Infection of Human HeLa Cells
Authors: Guerra Susana , López-Fernández Luis A. , Pascual-Montano Alberto , Muñoz Manuel , Harshman Keith , Esteban Mariano
Published: May 2003

Vaccinia virus (VV) is a cytocidal virus that causes major changes in host cell machinery shortly after infecting cells. To define the consequences of virus infection on host gene expression, we used microarrays of approximately 15,000 human cDNAs to examine expression levels of mRNAs isolated at 2, 6, and 16 h postinfection from cultures of infected HeLa cells. The majority of profiling changes during VV infection corresponded to downregulation of genes at 16 h postinfection. Differentially expressed genes were clustered into seven groups to identify common regulatory pathways, with most of them (90%) belonging to clusters 6 and 7, which represent genes whose expression was repressed after infection. Cluster 1, however, contained 37 transcripts (2.81%) showing a robust pattern of induction that was maintained during the course of infection. Genes in cluster 1 included those for Wiskott-Aldrich syndrome protein (WASP) family member WASF1, thymosine, adenosine A2a receptor, glutamate decarboxylase 2, CD-80 antigen, KIAA0888 protein, selenophosphate synthetase, pericentrin, and attractin as well as several expressed sequence tags. We analyzed in more detail the fate of WASP protein in VV-infected cells, because a related family member, N-WASP, is involved in viral motility. WASP protein accumulated in the course of infection; its increase required viral DNA replication and de novo protein synthesis, and it localized in cytoplasmic structures distinct from uninfected cells. This study is the first quantitative analysis of host gene expression following VV infection of cultured human cells, demonstrating global changes in the expression profile, and identifies upregulated genes with potential roles in the virus replication cycle.
Activation of Fusion by the SER Virus F Protein: a Low-pH-Dependent Paramyxovirus Entry Process
Authors: Seth Shaguna , Vincent Annelet , Compans R. W.
Published: May 2003

SER virus, a paramyxovirus closely related to simian virus 5, induces no syncytium formation. The SER virus F protein has a long cytoplasmic tail (CT), and truncation or mutations of the CT result in enhanced syncytium formation (S. Seth, A. Vincent, and R. W. Compans, J. Virol. 77:167-178, 2003; S. Tong, M. Li, A. Vincent, R. W. Compans, E. Fritsch, R. Beier, C. Klenk, M. Ohuchi, and H.-D. Klenk, Virology 301:322-333, 2002). We hypothesized that the presence of the long CT serves to stabilize the metastable conformation of the F protein. We observed that the hemifusion, cytoplasmic content mixing, and syncytium formation ability of the wild-type SER virus F coexpressed with the SER virus hemagglutinin-neuraminidase (HN) protein was enhanced, both qualitatively and quantitatively, at elevated temperatures. We also observed enhanced hemifusion, content mixing, and syncytium formation in SER virus F- and HN-expressing cells at reduced pH conditions ranging between 4.8 and 6.2. We have obtained evidence that in contrast to other paramyxoviruses, entry of SER virus into cells occurs by a low-pH-dependent process, indicating that the conversion to the fusion-active state for SER virus F is triggered by exposure to reduced pH.
Mechanisms of oxidative stress and vascular dysfunction
Authors: Nedeljkovic Z S , Gokce N , Loscalzo J
Published: May 2003

The endothelium regulates vascular homoeostasis through local elaboration of mediators that modulate vascular tone, platelet adhesion, inflammation, fibrinolysis, and vascular growth. Impaired vascular function contributes to the pathogenesis of atherosclerosis and acute coronary syndromes. There is growing pathophysiological evidence that increased generation of reactive oxygen species and oxidative stress participates in proatherogenic mechanisms of vascular dysfunction and atherothrombosis. In this review, the role of oxidative stress in mechanisms of vascular dysfunction is discussed, and potential antioxidant strategies are reviewed.
$K_L\to\gamma\nu\bar\nu$ DECAY BEYOND THE STANDARD MODEL
Authors: JIANG JI-HAO , GAO DAO-NENG , YAN MU-LIN
Published: May 2003

The decay [Formula: see text] is investigated beyond the standard model. Interestingly, the upper limit of the CP-conserving and CP-violating branching ratios of the decay, induced from the possible extensions of the standard model, would be larger than the corresponding branching ratios given in the standard model respectively, and it is expected that the CP-violating part could be enhanced.
Protein Phosphatases in Plants
Authors: Luan Sheng
Published: May 2003

Phosphorylation and dephosphorylation of a protein often serve as an “on-and-off” switch in the regulation of cellular activities. Recent studies demonstrate the involvement of protein phosphorylation in almost all signaling pathways in plants. A significant portion of the sequenced Arabidopsis genome encodes protein kinases and protein phosphatases that catalyze reversible phosphorylation. For optimal regulation, kinases and phosphatases must strike a balance in any given cell. Only a very small fraction of the thousands of protein kinases and phosphatases in plants has been studied experimentally. Nevertheless, the available results have demonstrated critical functions for these enzymes in plant growth and development. While serine/threonine phosphorylation is widely accepted as a predominant modification of plant proteins, the function of tyrosine phosphorylation, despite its overwhelming importance in animal systems, had been largely neglected until recently when tyrosine phosphatases (PTPs) were characterized from plants. This review focuses on the structure, regulation, and function of protein phosphatases in higher plants.
Molecular Mechanisms and Regulation of K+ Transport in Higher Plants
Authors: Véry Anne-Aliénor , Sentenac Hervé
Published: May 2003

Potassium (K+) plays a number of important roles in plant growth and development. Over the past few years, molecular approaches associated with electrophysiological analyses have greatly advanced our understanding of K+ transport in plants. A large number of genes encoding K+ transport systems have been identified, revealing a high level of complexity. Characterization of some transport systems is providing exciting information at the molecular level on functions such as root K+ uptake and secretion into the xylem sap, K+ transport in guard cells, or K+ influx into growing pollen tubes. In this review, we take stock of this recent molecular information. The main families of plant K+ transport systems (Shaker and KCO channels, KUP/HAK/KT and HKT transporters) are described, along with molecular data on how these systems are regulated. Finally, we discuss a few physiological questions on which molecular studies have shed new light.
Analyses of Polymorphism for UGT1*1 Exon 1 Promoter in Neonates with Pathologic and Prolonged Jaundice
Authors: Ülgenalp A. , Duman N. , Schaefer F.V. , Whetsell L. , Bora E. , Gülcan H. , Kumral A. , Ören H. , Giray Ö. , Erçal D. , Özkan H.
Published: May 2003

In this study, we investigated whether a TATA box polymorphism in the promoter of the UGT1*1 exon I, the most common detected DNA polymorphism in Gilbert’s syndrome, is a contributory factor in unexplained pathologic or prolonged jaundice. 38 neonates who had unexplained pathologic jaundice, 37 neonates who had unexplained prolonged jaundice, and 35 healthy, nonjaundiced neonates were enrolled in the study. Genotypes were assigned as follows: 6/6 (homozygous for a normal allele bearing the sequence [TA]6TAA), 7/7 (homozygous for an abnormal allele with the sequence [TA]7TAA), and 6/7 (heterozygous with one of each allele). Of the 110 infants, 10 (9%) had 7/7, 51 (46%) had 6/7, and 49 (45%) had 6/6 genotype; the differences between the three groups were not statistically significant. Also no differences were observed among different genotypes and mean serum total bilirubin concentrations. In conclusion, we showed that TA 7/7 and TA 6/7 genotypes are not rare in our population and that the presence of these polymorphisms alone does not play a significant role in the etiology of unexplained pathologic or prolonged neonatal hyperbilirubinemia.
Long Chain Fatty Acid Uptake by Human Intestinal Mucosa in vitro: Mechanisms of Transport
Authors: Chaves Célia Regina M. , Elias Paulo Roberto P. , Cheng Wanli , Zaltman Cyrla , Iglesias Antônio Carlos R. , Braulio Valeria B.
Published: May 2003

Background/Aims: The aim of this study was to investigate whether a carrier-mediated membrane transport for long chain fatty acid (LCFA) exists in the human intestinal mucosa. Methods: Biopsies obtained from the small intestine of patients, during routine endoscopies, were incubated for different periods in the presence of [3H]oleic acid complexed to albumin, and lipid fractions were separated and quantified. Biopsies were also incubated during 5 min in different oleic acid:albumin molar ratios. The effect of temperature on lipid transport was tested by incubating biopsies at 37 and 4°C. Results: Maximum [3H]oleic acid uptake occurred at 15 min. With increasing unbound oleic acid concentrations in the medium, LCFA uptake during 5 min of incubation revealed a typical saturation kinetics profile, and exhibiting apparent Km values of 3.43 ± 0.83 µM, and apparent Vmax values of 0.86 ± 0.07 nmol·min–1·mg–1. Within this period, nearly all intracellular fatty acid remained in the unesterified form. The system was also temperature-dependent, having shown a significant reduction of 78.03% in [3H]oleic acid uptake at 4°C, in 5 min of incubation. Conclusions: Our data confirm that LCFA uptake by human intestinal mucosa is likely to occur via a saturable carrier.
Effect of Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide on Pancreatic, Hepatic and Duodenal Mucosal Bicarbonate Secretion in the Pig
Authors: Glad Henrik , Ainsworth Mark A. , Svendsen Per , Fahrenkrug Jan , Schaffalitzky de Muckadell Ove B.
Published: May 2003

Background and Method: The potency of pituitary adenylate cyclase-activating polypeptide (PACAP) and the related vasoactive intestinal peptide (VIP) on pancreaticobiliary and duodenal mucosal bicarbonate secretion (DMBS) was studied in anaesthetized pigs. VIP, PACAP27 and PACAP38 were infused at doses of 250, 500, 1,000, 2,000 and 6,000 pmol/kg·h. Results: Infusion of VIP in these concentrations increased the portal plasma concentrations of VIP from 9 to 102, 220, 600, 850 and 3,795 pM, respectively. At doses of 1,000 and 6,000 pmol/kg·h all three peptides significantly stimulated pancreatic bicarbonate secretion, VIP from 0.0 to 0.3 and 7.6 mmol/h, PACAP27 from 0.0 to 2.3 and 8.2 mmol/h, and PACAP38 from 0.0 to 0.2 and 7.4 mmol/h, respectively. Only during infusion of 6,000 pmol/kg·h did the three peptides elicit a significant increase in hepatic bicarbonate output. At this dose VIP increased hepatic bicarbonate output from 0.4 to 3.8 mmol/h, PACAP27 from 0.3 to 3.6 mmol/h, and PACAP38 from 0.1 to 1.4 mmol/h, respectively. Infusion of VIP (500, 1,000, 2,000 and 6,000 pmol/kg·h) dose-dependently and significantly enlarged DMBS from 0.09 to 0.19, 0.41, 0.54 and 0.53 mmol/h, respectively. A significant effect of PACAP27 and PACAP38 on DMBS was only obtained at a dose of 6,000 pmol/kg·h, and at this dose VIP-induced DMBS was 4- and 8-fold higher than that induced by PACAP27 and PACAP38, respectively. Conclusions: VIP, PACAP27 and PACAP38 all have a pharmacological effect on both DMBS and pancreaticobiliary bicarbonate secretion. When comparing these results with the distribution of VIP/PACAP neurons and relevant receptors, it seems likely that VIP and PACAP have a physiological importance as neurotransmitters in the pancreas and duodenum, but not in the liver. The differences in the effect of VIP and PACAP on DMBS observed in this study indicate that there are variations between pancreatic and duodenal PACAP/VIP receptor subtypes, or that the duodenal inactivation of the peptides is different from the inactivation in the pancreas.
Biological Relevance of Terpenoids
Authors: Wagner Karl-Heinz , Elmadfa Ibrahim
Published: May 2003

The role of fruit and vegetables in human nutrition and public health are taken into account in most nutritional recommendations. Fruit and vegetables contain an abundance of phenolic substances, terpenoids and other natural antioxidants that have been associated with protection from and treatment of chronic diseases such as heart disease or cancer. Terpenoids are a group of substances which occur in nearly every natural food. Their main subclasses discussed as beneficial to maintain and improve health are monoterpenes (like limonene, carvone or carveol), diterpenes (including the retinoids), and tetraterpenes (which include all different carotenoids like α- and β-carotene, lutein, lycopene, zeaxanthine and cryptoxanthine). To be discussed as health promoting or biofunctional, the significant impact of a substance either on human metabolism or on well-defined and appropriate biomarkers must be shown. Based on the latter point, this paper reviews the literature on mono-, di- and tetraterpenes, with special focus on their impact on human health to answer the question of their biofunctionality. Special emphasis will be placed on their different mode of action, e.g. to affect oxidative stress, carcinogenesis and cardiovascular diseases.
Laboratory Tests in the Diagnosis of Pulmonary Embolism
Authors: Meyer Guy , Roy Pierre Marie , Sors Hervé , Sanchez Olivier
Published: May 2003

The diagnosis of pulmonary embolism (PE) requires objective testing. However, all imaging techniques have their own limitations and costs and cannot be performed in every patient with suspected PE. After decades of unfruitful research, several laboratory tests have been evaluated for suspected PE, the most promising being the D-dimer test. As a general rule, the specificity of D-dimers is too low to confirm PE. Conversely, several (but not all) D-dimer assays have a high sensitivity for diagnosing PE. Outcome studies indicate that the Vidas D-dimer and SimpliRED D-dimer can be used safely to withdraw anticoagulation when the pretest probability of PE is low (SimpliRED) or when it is low or moderate (Vidas). These results may however not apply to other D-dimer assays and clinicians should know the characteristics of the test used in their hospital. Blood gas analysis does not have sufficient sensitivity and specificity to confirm or exclude PE, but it may be used to evaluate the clinical probability of PE before other testing is done. The diagnostic value of the alveolar dead space fraction in patients with suspected PE is currently investigated. Initial data suggest that it needs to be combined with a D-dimer test to safely exclude PE. Brain natriuretic peptide and cardiac troponin have limited usefulness for diagnosing PE, but both tests may identify patients with a poor prognosis, in whom more aggressive treatment may be warranted.
Bilateral and Unilateral Spontaneous Massive Hemothorax as a Presenting Manifestation of Rare Tumors
Authors: Varsano Shabtai , Edelstein Evgeny , Gendel Boris , Smorzik Joshua
Published: May 2003

Spontaneous true hemothorax is quite a rare manifestation of a presenting disease. This is a report of two patients, one with bilateral spontaneous massive hemothorax as a presenting manifestation of angiosarcoma involving the lungs and pleura, and the other with unilateral spontaneous hemothorax and hemorrhagic shock as a presenting manifestation of ‘cystic’ chondroblastoma. Differential diagnosis of spontaneous true hemothorax and its evaluation and management are discussed.
Experimental Trigeminal Glycerol Injection in Dogs: Histopathological Evaluation by Light and Electron Microscopy
Authors: Işik N. , Pamir M.N. , Benli K. , Erbengi A. , Erbengi T. , Ruacan Ş.
Published: May 2003

We investigated the effects of percutaneous gasserian glycerol injection in dogs and reviewed the histopathological changes. Experiments were performed in 16 adult healthy mongrel dogs. In group 1 (8 dogs) normal saline and in group 2 (8 dogs) pure glycerol was injected in the right trigeminal ganglion. After these procedures, dogs in each group were sacrificed after 24 h (3 dogs), 7 days (3 dogs), 21 days (2 dogs). The trigeminal ganglion and nerve of both sides were removed by using microsurgical techniques and examined by light and electron microscopy. Group 1: in all sections, nerve cells, myelinated and nonmyelinated fibers revealed normal patterns with slight fibrosis. Group 2: in all sections, myelinated fibers showed disintegration and swelling of the myelin sheath, rupture of axon continuity, destruction of basal lamina, deformation of the myelin-axon relationship by both light microscopy and electron microscopy. The sections examined by electron microscopy also showed axonolysis in nonmyelinated fibers. The changes after 7 and 21 days were less prominent than after 24 h. In the left sides, there are no pathological changes. Glycerol has a neurolytic effect on the dog’s trigeminal ganglion. These effects were not specific and selective for myelinated and nonmyelinated nerve fibers.
Management and Follow-Up of Impacted Ureteral Stones
Authors: Deliveliotis C. , Chrisofos M. , Albanis S. , Serafetinides E. , Varkarakis J. , Protogerou V.
Published: May 2003

Introduction: Impacted stones are those that remain unchanged in the same location for at least 2 months. Materials and Methods: We evaluated 42 patients with impacted ureteral stones, and followed them for two and a half years to check for long-term results. The calculi location included all three segments of the ureter (proximal, mid and distal). Patients’ age ranged from 22 to 83 years (mean 52.5 years). Primarily, patients were manipulated with extracorporeal shock wave lithotripsy (ESWL) in situ, or following stenting. If the result was not satisfactory, then we proceeded to retrograde ureteroscopy and ureterolithotripsy. Open ureterolithotomy was our final choice. Results: Thirty-six of the 42 patients (85.7%) were stone-free without the need of an open procedure. Follow-up period ranged from 10 up to 40 months, with a median period of 30 months and was achieved in 30 patients (71.4%). Stone recurrence was noted in 4 cases, while hydronephrosis without evidence of stone presence in 2. Conclusions: The initial approach for the treatment of impacted lithiasis should be attempted by ESWL. If this fails, alternative therapeutic solutions such as endoscopy can result in removal of the stone.
Association of E-Cadherin Gene 3’-UTR C/T Polymorphism with Calcium Oxalate Stone Disease
Authors: Tsai Fuu-Jen , Wu Hsi-Chin , Chen Huey-Yi , Lu Hsueh-Fu , Hsu Cheng-Der , Chen Wen-Chi
Published: May 2003

Introduction: Urinary stone disease is one of the most commonly seen urological diseases in Taiwan. Single nucleotide polymorphisms (SNPs) are commonly used for the investigation of genetic markers for stone disease. E-cadherin (CDH-1) is one of the cellular junction proteins related to the integrity of epithelial cells. Our aim was to investigate a polymorphism of the CDH-1 gene 3′-UTR as a possible genetic marker in the search for the genetic causes. Materials and Methods: 148 patients with calcium oxalate stone were compared with 103 healthy controls for the frequency of CDH-1 3′-UTR polymorphisms. The polymorphism was detected by polymerase chain reaction-based restriction analysis (Pml I endonuclease). Results and Conclusions: The results revealed significant differences between normal individuals and calcium stone disease patients (p = 0.0013). The distribution of genotype TT homozygote was higher in stone patients (51.5%) than in the control group (43.4%). The odds ratio for T allele compared to C allele was 2.0. We have concluded that polymorphisms of CDH-1 3′-UTR is a valid genetic marker for calcium stone disease.
3-Month Formulation of Goserelin Acetate (‘Zoladex’ 10.8-mg Depot) in Advanced Prostate Cancer: Results from an Italian, Open, Multicenter Trial
Authors: Fontana D. , Mari M. , Martinelli A. , Boccafoschi C. , Magno C. , Turriziani M. , Maymone S.S. , Cunico S. Cosciani , Zanollo A. , Montagna G. , Frongia M. , Jacobellis U.
Published: May 2003

Objectives: To determine the endocrine effects, efficacy and tolerability of the 3-month formulation of goserelin acetate (‘Zoladex’ 10.8-mg depot; ‘Zoladex’ is a trade mark of the AstraZeneca group of companies) in the treatment of patients with advanced prostate cancer. Methods: Between February 1996 and October 1997, this open, multicentre study enrolled 120 patients with locally advanced (T3/4) or metastatic (N+ or M1) disease, or an increase in prostate-specific antigen (PSA) level after radical prostatectomy. Patients received goserelin acetate 10.8-mg depot every 12 weeks until clinical progression or interruption for adverse events or other reasons. Results: The mean testosterone concentrations were suppressed to the castration range (≤2 nmol/l) after 4 weeks of treatment and remained suppressed throughout the study. In total, 99/115 (86%) patients had a serum PSA response, and the mean PSA value decreased significantly during treatment (p = 0.006). The mean PSA level at baseline was significantly lower in patients without disease progression compared to those who experienced disease progression (p = 0.0002). Goserelin acetate 10.8-mg depot was well tolerated and there were no injection site reactions. Conclusions: The goserelin acetate 10.8-mg depot is well tolerated with no injection site reactions. It produces PSA responses and provides reliable suppression of serum testosterone.
Clinical Trials: Evidence and Unanswered Questions – Hypertension
Authors: Celermajer David S.
Published: May 2003

Since the pioneering publications of the Hypertension Detection and Follow-up Program (HDFP) and the Multiple Risk Factor Intervention Trial (MRFIT) in the late 1970s and early 1980s, it has become established that lowering blood pressure in high-risk patients is a highly effective form of primary prevention for stroke. Over the subsequent 25 years, over 30 large clinical trials have extended these initial observations to allow us to conclude that treatment of mild, moderate or severe hypertension, and isolated systolic hypertension in the elderly, all produce important absolute benefits. In addition, excellent specific evidence of benefit is now accumulating for certain groups of normotensive patients, including those with previous stroke, and those with established cardiovascular disease. Although the importance of vigorous antihypertensive therapy for the primary and secondary prevention of stroke is increasingly clear, a large number of unanswered questions remain. For example, while it is apparent that diuretics, β-blockers, calcium channel blockers (CCBs) and angiotensin-converting enzyme (ACE) inhibitors are all effective antihypertensive agents, the question remains as to which drug, or combination of drugs, is best for which patients. The results of several ongoing comparative trials of different drug regimens, including the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT), may elucidate this further.
Sirolimus (Rapamycin) Monotherapy Prevents Graft Vascular Disease in Nonhuman Primate Recipients of Orthotopic Aortic Allografts
Authors: Dambrin Camille , Klupp Jochen , Bîrsan Tudor , Luna Jorge , Suzuki Takeshi , Lam Tuan , Stähr Peter , Hausen Bernard , Christians Uwe , Fitzgerald Peter , Berry Gerald , Morris Randall
Published: May 2003

Background—

Delayed treatment with sirolimus (SRL) halts progression of graft vascular disease (GVD) in nonhuman primate (NHP) aortic allograft recipients. In this study, we investigated whether SRL monotherapy prevents the development of GVD.

Methods and Results—

Pairs of 3-cm infrarenal aortic segments were exchanged between mixed lymphocyte reaction–mismatched, blood group–compatible NHPs (n=12). Six NHPs were untreated controls, and 6 were treated orally with SRL starting on the day of transplantation. Follow-up was 105 days. SRL doses were adjusted individually by assessing SRL blood concentrations, immune function, and clinical status. The severity of GVD was determined every 3 weeks by intravascular ultrasound, which quantified intimal area (IA) and intimal volume (IV) for the middle 1-cm graft segments. The mean±SEM SRL plasma levels were 14.5±9 ng/mL. In grafts from treated NHPs, IA and IV values on days 63, 84, and 105 were significantly lower than for controls (
P
<0.05 to
P
<0.001). On day 105, in the grafts from SRL-treated NHPs compared with grafts from controls, values (mean±SEM) were IA, 2.9±0.9 versus 5.5±0.7 mm
2
,
P
<0.001 and IV, 29.6±4.6 versus 55.2±2.8 mm
3
,
P
<0.001; IA and IV values for grafts from SRL-treated NHPs did not increase significantly between days 21 and 105.

Conclusions—

We show that SRL monotherapy prevented GVD in NHP aortic allograft recipients, suggesting the value of SRL for controlling GVD in clinical transplantation.
Multiply Attenuated, Self-Inactivating Lentiviral Vectors Efficiently Deliver and Express Genes for Extended Periods of Time in Adult Rat Cardiomyocytes In Vivo
Authors: Fleury Sylvain , Simeoni Eleonora , Zuppinger Christian , Déglon Nicole , von Segesser Ludwig K. , Kappenberger Lukas , Vassalli Giuseppe
Published: May 2003

Background—

Among retroviral vectors, lentiviral vectors are unique in that they transduce genes into both dividing and nondividing cells. However, their ability to provide sustained myocardial transgene expression has not been evaluated.

Methods and Results—

Multiply attenuated, self-inactivating lentivectors based on human immunodeficiency virus-1 contained the enhanced green fluorescent protein (EGFP) gene under the transcriptional control of either the cytomegalovirus (CMV) immediate-early enhancer/promoter, the elongation factor-1α (EF-1α) promoter, or the phosphoglycerate-kinase (PGK) promoter. Lentivectors transduced adult rat cardiomyocytes in a dose-dependent manner (transduction rates, >90%; multiplicity of infection, ≈5). The CMV promoter achieved higher EGFP expression levels than the EF-1α and PGK promoters. Insertion of the central polypurine tract
pol
sequence improved gene transfer efficiency by ≈2-fold. In vivo gene transfer kinetics was studied by measuring the copy number of integrated lentivirus DNA and EGFP concentrations in cardiac extracts by real-time polymerase chain reaction and ELISA, respectively. With CMV promoter-containing lentivectors, vector DNA peaked at day 3, declined by ≈4-fold at day 14, but then remained stable up to week 10. Similarly, EGFP expression peaked at day 7, decreased by ≈7-fold at day 14, but was essentially stable thereafter. In contrast, vector DNA and EGFP expression declined rapidly with EF-1α promoter–containing lentivectors. Peak EGFP expression with titer-matched adenovectors was ≈35% higher than with CMV lentivectors but was lost rapidly over time.

Conclusions—

Lentivectors efficiently transduce and express genes for extended periods of time in cardiomyocytes in vivo. Lentivectors provide a useful tool for studying myocardial biology and a potential system for gene heart therapy.
L-4F, an Apolipoprotein A-1 Mimetic, Dramatically Improves Vasodilation in Hypercholesterolemia and Sickle Cell Disease
Authors: Ou Jingsong , Ou Zhijun , Jones Deron W. , Holzhauer Sandra , Hatoum Ossama A. , Ackerman Allan W. , Weihrauch Dorothee W. , Gutterman David D. , Guice Karen , Oldham Keith T. , Hillery Cheryl A. , Pritchard Kirkwood A.
Published: May 2003

Background—

Hypercholesterolemia and sickle cell disease (SCD) impair endothelium-dependent vasodilation by dissimilar mechanisms. Hypercholesterolemia impairs vasodilation by a low-density lipoprotein (LDL)–dependent mechanism. SCD has been characterized as a chronic state of inflammation in which xanthine oxidase (XO) from ischemic tissues increases vascular superoxide anion (O
2
·
−
) generation. Recent reports indicate that apolipoprotein (apo) A-1 mimetics inhibit atherosclerosis in LDL receptor–null (
Ldlr
−/−
) mice fed Western diets. Here we hypothesize that L-4F, an apoA-1 mimetic, preserves vasodilation in hypercholesterolemia and SCD by decreasing mechanisms that increase O
2
·
−
generation.

Methods and Results—

Arterioles were isolated from hypercholesterolemic
Ldlr
−/−
mice and from SCD mice that were treated with either saline or L-4F (1 mg/kg per day). Vasodilation in response to acetylcholine was determined by videomicroscopy. Effects of L-4F on LDL-induced increases in endothelium-dependent O
2
·
−
generation were determined on arterial segments via the hydroethidine assay and on stimulated endothelial cell cultures via superoxide dismutase–inhibitable ferricytochrome
c
reduction. Effects of L-4F on XO bound to pulmonary arterioles and content in livers of SCD mice were determined by immunofluorescence. Hypercholesterolemia impaired vasodilation in
Ldlr
−/−
mice, which L-4F dramatically improved. L-4F inhibited LDL-induced increases in O
2
·
−
in arterial segments and in stimulated cultures. SCD impaired vasodilation, increased XO bound to pulmonary endothelium, and decreased liver XO content. L-4F dramatically improved vasodilation, decreased XO bound to pulmonary endothelium, and increased liver XO content compared with levels in untreated SCD mice.

Conclusions—

These data show that L-4F protects endothelium-dependent vasodilation in hypercholesterolemia and SCD. Our findings suggest that L-4F restores vascular endothelial function in diverse models of disease and may be applicable to treating a variety of vascular diseases.
Consolidation Docetaxel After Concurrent Chemoradiotherapy in Stage IIIB Non–Small-Cell Lung Cancer: Phase II Southwest Oncology Group Study S9504
Authors: Gandara David R. , Chansky Kari , Albain Kathy S. , Leigh Bryan R. , Gaspar Laurie E. , Lara Primo N. , Burris Howard , Gumerlock Paul , Kuebler J. Philip , Bearden James D. , Crowley John , Livingston Robert
Published: May 2003

Purpose: To test the concept of taxane sequencing in combined-modality therapy, this phase II trial (S9504) evaluated consolidation docetaxel after concurrent chemoradiotherapy in patients with pathologically documented stage IIIB non–small-cell lung cancer (NSCLC). Results were compared with those of the predecessor study (S9019) with identical eligibility, staging criteria, and treatment, excepting docetaxel consolidation.

Patients and Methods: Treatment consisted of cisplatin 50 mg/m2 on days 1, 8, 29, and 36, etoposide 50 mg/m2 on days 1 through 5 and 29 through 33, and concurrent thoracic radiotherapy (total dose of 61 Gy). Consolidation docetaxel started 4 to 6 weeks after chemoradiotherapy at an initial dose of 75 mg/m2.

Results: Stage subsets (tumor-node-metastasis system) in 83 eligible patients were as follows: T4N0/1, 31 patients (37%); T4N2, 22 patients (27%), and T1–3N3, 30 patients (36%). Concurrent chemoradiotherapy was generally well tolerated, but two patients died from probable radiation-associated pneumonitis. Neutropenia during consolidation docetaxel was common (57% with grade 4) and most frequent during escalation to 100 mg/m2. Median progression-free survival was 16 months, median survival was 26 months, and 1-, 2-, and 3-year survival rates were 76%, 54%, and 37%, respectively. Brain metastasis was the most common site of failure. In S9019, median survival was 15 months and 1-, 2-, and 3-year survival rates were 58%, 34%, and 17%, respectively.

Conclusion: Consolidation docetaxel after concurrent chemoradiotherapy in stage IIIB NSCLC is feasible and generally tolerable, and results compare favorably with the predecessor trial S9019. Nevertheless, this study remains hypothesis-generating and does not provide definitive evidence of the benefit of this approach. Phase III trials evaluating the S9504 regimen have been initiated to validate these results.
Molecular Grading of Urothelial Cell Carcinoma With Fibroblast Growth Factor Receptor 3 and MIB-1 is Superior to Pathologic Grade for the Prediction of Clinical Outcome
Authors: van Rhijn Bas W.G. , Vis André N. , van der Kwast Theo H. , Kirkels Wim J. , Radvanyi François , Ooms Engelbert C.M. , Chopin Dominique K. , Boevé Egbert R. , Jöbsis Adriaan C. , Zwarthoff Ellen C.
Published: May 2003

Purpose: Fibroblast growth factor receptor 3 (FGFR3) mutations were recently found at a high frequency in well-differentiated urothelial cell carcinoma (UCC). We investigated the relationship between FGFR3 status and three molecular markers (MIB-1, P53, and P27kip1) associated with worse prognosis and determined the reproducibility of pathologic grade and molecular variables.

Patients and Methods: In this multicenter study, we included 286 patients with primary (first diagnosis) UCC. The histologic slides were reviewed. FGFR3 status was examined by polymerase chain reaction–single strand conformation polymorphism and sequencing. Expression levels of MIB-1, P53, and P27kip1 were determined by immunohistochemistry. Mean follow-up was 5.5 years (range, 0.4 to 18.4 years).

Results: FGFR3 mutations were detected in 172 (60%) of 286 UCCs. Grade 1 tumors had an FGFR3 mutation in 88% of patient samples and grade 3 tumors in 16% of patient samples. Conversely, aberrant expression patterns of MIB-1, P53, and P27kip1 were seen in 5%, 2%, and 3% of grade 1 tumors and in 85%, 60%, and 56% of grade 3 tumors, respectively. In multivariate analysis with recurrence rate, progression, and disease-specific survival as end points, the combination of FGFR3 and MIB-1 proved independently significant in all three cases. By using these two molecular markers, three molecular grades (mGs) could be identified: mG1 (mutation; normal expression), favorable prognosis; mG2 (two remaining combinations), intermediate prognosis; and mG3 (no mutation; high expression), poor prognosis. The molecular variables were more reproducible than pathologic grade (85% to 100% v 47% to 61%).

Conclusion: The FGFR3 mutation represents the favorable molecular pathway of UCC. Molecular grading provides a new, simple, and highly reproducible tool for clinical decision making in UCC patients.
Quality-of-Care Indicators for Early-Stage Prostate Cancer
Authors: Spencer Benjamin A. , Steinberg Michael , Malin Jennifer , Adams John , Litwin Mark S.
Published: May 2003

Purpose: Decisions regarding treatment for early-stage prostate cancer are frustrated not only by inadequate evidence favoring one treatment modality but also by the absence of data comparing quality among providers. In fact, the choice of provider may be as important as the choice of treatment. We undertook this study to develop an infrastructure to evaluate variations in quality of care for men with early-stage prostate cancer.

Methods: We enlisted several sources to develop a list of proposed quality-of-care indicators and covariates. After an extensive structured literature review and a series of focus groups with patients and their spouses, we conducted structured interviews with national academic leaders in prostate cancer treatment. We then convened an expert panel using the RAND consensus method to discuss and rate the validity and feasibility of the proposed quality indicators and covariates.

Results: The panel endorsed 49 quality-of-care indicators and 14 covariates, which make up our final list of candidate measures. Several domains of quality are represented in the selected indicators, including patient volume, pretreatment referrals, preoperative testing, interpretation of pathology specimens, and 10-year disease-free survival. Covariates include measures of case-mix, such as patient age and comorbidity.

Conclusion: This study establishes a foundation on which to build quality-of-care assessment tools to evaluate the treatment of early-stage prostate cancer. The next step is to field-test the indicators for feasibility, reliability, validity, and clinical utility in a population-based sample. This work will begin to inform medical decision-making for patients and their physicians.
Role of Telomeres and Telomerase in the Pathogenesis of Human Cancer
Authors: Hahn William C.
Published: May 2003

Specialized nucleoprotein structures, termed telomeres, cap the ends of human chromosomes. These terminal structures, composed of repetitive arrays of guanine-rich hexameric DNA together with specific telomere-binding proteins, play essential roles in protecting the chromosome from damage and degradation. In addition, several lines of evidence implicate telomere maintenance as an important regulator of cell life span. Activation of telomerase, a dedicated reverse transcriptase that synthesizes telomeric sequences, is strongly associated with cancer, and recent observations confirm that telomeres and telomerase perform important roles in both suppressing and facilitating malignant transformation. These dual functions of telomere biology are evident in the clinical manifestations of the multisystem syndrome, dyskeratosis congenita, forms of which display defects in telomerase function. Recent advances in our understanding of telomere biology indicate that the manipulation of telomeres and telomerase will lead to clinically significant applications in the diagnosis, prevention, and treatment of neoplastic disease.
Adiposity and Sex Hormones in Postmenopausal Breast Cancer Survivors
Authors: McTiernan Anne , Rajan Kumar B. , Tworoger Shelley S. , Irwin Melinda , Bernstein Leslie , Baumgartner Richard , Gilliland Frank , Stanczyk Frank Z. , Yasui Yutaka , Ballard-Barbash Rachel
Published: May 2003

Purpose: Overweight and obese women with breast cancer have poorer survival compared with thinner women. One possible reason is that breast cancer survivors with higher degrees of adiposity have higher concentrations of tumor-promoting hormones. This study examined the association between adiposity and concentrations of estrogens, androgens, and sex hormone–binding globulin (SHBG) in a population-based sample of postmenopausal women with breast cancer.

Methods: We studied the associations between body mass index (BMI), body fat mass, and percent body fat, measured by dual-energy x-ray absorptiometry scan, waist circumference, and waist-to-hip circumference ratio, with concentrations of estrone, estradiol, testosterone, SHBG, dehydroepiandrosterone sulfate, free estradiol, and free testosterone in 505 postmenopausal women in western Washington and New Mexico with incident stage 0 to IIIA breast cancer. Blood and adiposity measurements were performed between 4 and 12 months after diagnosis.

Results: Obese women (BMI ≥ 30) had 35% higher concentrations of estrone and 130% higher concentrations of estradiol compared with lighter-weight women (BMI < 22.0; P = .005 and .002, respectively). Similar associations were observed for body fat mass, percent body fat, and waist circumference. Testosterone concentrations also increased with increasing levels of adiposity (P = .0001). Concentrations of free estradiol and free testosterone were two to three times greater in overweight and obese women compared with lighter-weight women (P = .0001).

Conclusion: These data provide information about potential hormonal explanations for the association between adiposity and breast cancer prognosis. These sex hormones may be useful biomarkers for weight loss intervention studies in women with breast cancer.
Combination of Fludarabine and Mitoxantrone in Untreated Stages III and IV Low-Grade Lymphoma: S9501
Authors: Velasquez William S. , Lew Danika , Grogan Thomas M. , Spiridonidis C. Harris , Balcerzak Stanley P. , Dakhil Shaker R. , Miller Thomas P. , Lanier Keith S. , Chapman Robert A. , Fisher Richard I.
Published: May 2003

Purpose: To determine the efficacy of combination fludarabine and mitoxantrone (FN) in untreated stages III and IV low-grade lymphoma. The major end point was to estimate progression-free survival (PFS) in all eligible patients.

Patients and Methods: Seventy-eight eligible patients were registered. Chemotherapy courses were administered every 4 weeks with mitoxantrone 10 mg/m2 on day 1 and fludarabine 25 mg/m2 on days 1, 2, and 3 for a total of six to eight cycles. Pneumocystis carinii prophylaxis was required.

Results: Seventy-three patients (94%) attained an objective response. Complete remission was demonstrated in 34 patients (44%) and partial remission was demonstrated in 39 patients (50%). With a median follow-up time of 5.5 years, the median PFS was 32 months, with a 4-year PFS rate of 38%. Median survival has not been reached and 88% of all patients are alive at 4 years. The application of the International Prognostic Index and serologic staging showed significant differences in PFS in all risk groups, whereas overall survival was markedly worse for the highest-risk group in either prognostic model. Three prior Southwest Oncology Group trials using a regimen of cyclophosphamide, doxorubicin, vincristine, and prednisone or a combination of prednisone, vincristine, methotrexate, cytarabine, cyclophosphamide, etoposide, nitrogen mustard, vincristine, procarbazine, and prednisone in similar patient populations demonstrated comparable clinical outcome, although the 4-year survival for FN was better. FN was well tolerated, but mild to severe reversible myelosuppression was noted. Other complications were rare.

Conclusion: FN is an effective, safe chemotherapy combination for patients with advanced-stage, low-grade lymphoma. Clinical outcomes were comparable to prior published data using anthracycline-based regimens.
Contribution of Variable-Speed Wind Turbines to Voltage Control
Authors: Ledesma Pablo , Usaola Julio
Published: May 2003

Variable speed, grid connected, wind turbines open new possibilities for voltage control, because they use electronic converters, which may regulate the reactive power interchange with the grid. This paper proposes two voltage control schemes for variable speed wind turbines with double-fed induction generator. The first scheme acts on the wind-turbine power factor, while the latter acts directly on the converter current. Advantages and drawbacks of both techniques are discussed. Both control techniques have been tested by simulations of a base case, which represent a synchronous generator, a wind farm and a local load, and several disturbances such as the loss of compensator capacitors.
Reliability and Safety Assessment of Wind Turbines Control and Protection Systems
Authors: Michos D. , Dialynas E. , Vionis P.
Published: May 2003

The paper describes a method for the reliability and safety assessment of wind turbine control and protection systems. This enables the qualitative and quantitative reliability analysis of these systems and calculates an appropriate set of reliability indices. The method for the probabilistic safety assessment of these systems includes the event-tree approach for identifying all possible unsafe events of wind turbines operation and all scenarios that can lead to them. The Markov procedure is used for the modelling of all failure modes. This method is compatible with the existing respective international safety standards and criteria; it can be used by designers, manufacturers and certification institutes. The analysis of a part of the control and protection system of the CRES wind turbine in Agios Efstratios is also included.
Comparison between Types of Generation for Wind Turbines Operating in Stochastic Wind
Authors: Kanellos F. D. , Hatziargyriou N. D.
Published: May 2003

This paper analyses the repercussions from the connection of Wind Parks on the operation of weak electricity distribution systems, based on simulation results. Three different cases concerning Fixed Speed Wind Turbines are studied. Also Variable Speed Wind Turbines are examined, using (i), doubly-fed induction machines, and (ii), a dual voltage-source converter cascade. All these types of wind turbines are assumed to be equipped with asynchronous machines. The effects on the voltage profile caused by both Fixed and Variable Speed Wind Turbines are compared. The advantages of the variable speed operation are confirmed. The possible increase in the installed capacity of a Wind Park with variable speed turbines, as compared with fixed speed turbines maintaining the same power quality standards, is estimated. The distribution system studied is an actual feeder located in a rural area at the southeast of mainland Greece. This area presents excellent wind potential and as a result independent power producers have expressed a considerable interest for the installation of Wind Parks.