bluuebunny/crossref_metadata_2025 huggingface dataset:
German Vegan Study: Diet, Life-Style Factors, and Cardiovascular Risk Profile
Authors: Waldmann A. , Koschizke J.W. , Leitzmann C. , Hahn A.
Published: October 2005

Background/Aim: Evaluation of cardiovascular risk profile in 154 German vegans. Methods: Cross-sectional study, Germany. Study instruments: 2 FFQ, 2 questionnaires, analyses of fasting venous blood samples. Results: The total study population had a low BMI (mean: 22.3 kg/m2), a moderate blood pressure (mean: 120/75 mm Hg), an extremely low consumption of alcohol (mean: 0.77 g/day) and 96.8% were nonsmokers. Moderate physical activity (PAL) was reported by nearly 50%, whereas 22.7% declared to have a high PAL (>3 h/week). Median triacylglycerol (TG) was 0.81 mmol/l, total cholesterol (TC) was 4.33 mmol/l, HDL was 1.34 mmol/l. The mean TC/HDL-ratio was 3.3. Lipoprotein(a) (Lp(a)) was 8.13 mg/dl, concentrations of >30 mg/dl were prevalent in 25% of the participants. In general, status of folate and pyridoxine were sufficient, while 49.7% showed cobalamin concentrations <150 pmol/l. Plasma homocysteine levels were slightly elevated (median: 12.5 µmol/l). Cobalamin concentration and duration of vegan nutrition were the main determinants of homocysteine in the total study population. Conclusion: Although TC and LDL concentrations were favorable, low HDL and elevated homocysteine and Lp(a) concentrations were unfavorable. Overall, these results confirm the notion that a vegan diet is deficient in vitamin B12, which may have an unfavorable effect on CHD risk.
A High-Cholesterol, n–3 Polyunsaturated Fatty Acid Diet Causes Different Responses in Rats and Hamsters
Authors: Lin Mei-Huei , Lu Shao-Chun , Huang Po-Chao , Liu Young-Chau , Liu Shyun-Yeu
Published: October 2005

This study was designed to investigate the response to a high-cholesterol, n–3 polyunsaturated fatty acid (PUFA) or n–6 PUFA diet in rats and hamsters. Animals were fed n–3 or n–6 PUFA with a cholesterol-free diet, or with a diet enriched with cholesterol (0.5%, w/w) for 2 weeks. In rats and hamsters fed a cholesterol-free diet, plasma cholesterol, triglycerides and very-low-density lipoprotein (VLDL)-triglyceride levels in n–3 PUFA group were significantly lower than those in n–6 PUFA group. In contrast, when diets were supplemented with 0.5% cholesterol, the plasma cholesterol- and triglyceride-lowering effect of dietary n–3 PUFA disappeared. In hamsters fed with the atherogenic diet (0.5% dietary cholesterol) for 2 weeks, n–3 PUFA induced hypercholesterolemia more than n–6 PUFA, the increase being in the VLDL and low-density lipoprotein (LDL) fractions. Our data thus indicate that elevation of VLDL- and LDL-cholesterol in hamsters by n–3 PUFA, compared with n–6 PUFA, is dependent on 0.5% dietary cholesterol supplementation. In rats, on the other hand, dietary n–3 PUFA did not induce hypercholesterolemia more than n–6 PUFA when 0.5% cholesterol was supplemented. Although the effects of n–3 PUFA on plasma cholesterol, triglycerides and VLDL-triglycerides were similar in hamsters and rats, the interactive effects of n–3 PUFA and cholesterol on plasma and lipoprotein cholesterol levels differed in the two species. It was also found that plasma triglycerides, cholesterol and lipoprotein cholesterol levels in hamsters are higher than in rats in the presence and absence of dietary cholesterol. In addition, cholesterol feeding induces hypertriglyceridemia and hypercholesterolemia only in hamsters. Moreover, liver triglyceride concentrations increased in rats fed a cholesterol-rich diet and hepatic triglyceride levels of the n–3 PUFA-fed rats were significantly lower than those in the n–6 PUFA-fed rats in the presence and absence of dietary cholesterol. However, triglycerides did not accumulate in the liver in hamsters fed a cholesterol-rich diet and hepatic triglyceride levels of the n–3 PUFA-fed hamsters were not significantly different from those in the n–6 PUFA-fed hamsters in the presence and absence of dietary cholesterol. Therefore, these studies confirm marked species differences in response to the interactive effects of dietary n–3 PUFA and cholesterol.
Intra- and Extrauterine Maturation of Amplitude-Integrated Electroencephalographic Activity in Preterm Infants Younger than 30 Weeks of Gestation
Authors: Klebermass Katrin , Kuhle Stefan , Olischar Monika , Rücklinger Ernst , Pollak Arnold , Weninger Manfred
Published: October 2005

Objective: To prospectively investigate the longitudinal changes of amplitude-integrated electroencephalographic (aEEG) activity in preterm infants <30 weeks gestational age (GA). Methods: Infants (GA <30 weeks) without evidence of neurological abnormalities had weekly aEEG recordings performed. The relative duration of the three aEEG patterns (discontinuous low voltage, discontinuous high voltage and continuous) was determined and the influence of GA and postnatal age (PNA) on the occurrence of each pattern was assessed. Results: Ninety-eight infants (median GA 26 weeks; range 23–29 weeks) were studied. With higher GA (OR 1.68, 95% CI 1.33–2.13) and PNA (OR 1.91, 95% CI 1.53–2.38), the likelihood for the occurrence of continuous activity increased. The discontinuous low-voltage pattern was less likely to occur with increasing GA (OR 0.68, 95% CI 0.55–0.83) and PNA (OR 0.70, 95% CI 0.61–0.81). Conclusion: Maturation of aEEG activity in preterm infants is influenced by both GA and PNA.
SimPed: A Simulation Program to Generate Haplotype and Genotype Data for Pedigree Structures
Authors: Leal Suzanne M. , Yan Kai , Müller-Myhsok Bertram
Published: October 2005

With the widespread availability of SNP genotype data, there is great interest in analyzing pedigree haplotype data. Intermarker linkage disequilibrium for microsatellite markers is usually low due to their physical distance; however, for dense maps of SNP markers, there can be strong linkage disequilibrium between marker loci. Linkage analysis (parametric and nonparametric) and family-based association studies are currently being carried out using dense maps of SNP marker loci. Monte Carlo methods are often used for both linkage and association studies; however, to date there are no programs available which can generate haplotype and/or genotype data consisting of a large number of loci for pedigree structures. SimPed is a program that quickly generates haplotype and/or genotype data for pedigrees of virtually any size and complexity. Marker data either in linkage disequilibrium or equilibrium can be generated for greater than 20,000 diallelic or multiallelic marker loci. Haplotypes and/or genotypes are generated for pedigree structures using specified genetic map distances and haplotype and/or allele frequencies. The simulated data generated by SimPed is useful for a variety of purposes, including evaluating methods that estimate haplotype frequencies for pedigree data, evaluating type I error due to intermarker linkage disequilibrium and estimating empirical p values for linkage and family-based association studies.
Is Alexithymia Related to Negative Symptoms of Schizophrenia?
Authors: Todarello Orlando , Porcelli Piero , Grilletti Francesco , Bellomo Antonello
Published: October 2005

Background: Alexithymic features are close to anhedonia, blunted affect, and alogia that are also characteristics of the negative symptoms of schizophrenia. This study aimed to evaluate whether alexithymia is associated with negative symptoms and is related to the change of schizophrenic symptoms over time. Sampling and Methods: A consecutive sample of 29 schizophrenic outpatients was evaluated at baseline and at 3, 6, and 12 months during appropriate treatment. They completed the Positive and Negative Syndrome Scale, the Montgomery and Asberg Depression Rating Scale, the Global Assessment of Functioning Scale, and the 20-item Toronto Alexithymia Scale at any time points. Results: The psychiatric scale scores showed significant symptom improvement over time but were unrelated to the alexithymia score that was instead stable over time. Hierarchical regression showed that the 20-item Toronto Alexithymia Scale at baseline was the sole predictor of alexithymia at 12 months, after controlling for psychopathology and psychological functioning. Conclusions: Alexithymia was unrelated to negative symptoms, suggesting it is an independent and separate construct from schizophrenia. As expected, the negative symptoms were associated instead with illness-related aspects of depression and psychosocial functioning. Caution should be expressed in generalization mainly because this study is limited by the small sample size.
Depressive and Anxiety Disorder Comorbidity in Obsessive Compulsive Disorder
Authors: Bhattacharyya Sagnik , Reddy Y.C. Janardhan , Khanna S.
Published: October 2005

Background: Depressive and other anxiety disorders are commonly found to coexist with obsessive compulsive disorder (OCD). Although western studies have looked at this issue, there are no reports from India investigating anxiety and depressive comorbidity in adult OCD. Methods: Between January and December 2001, charts of 218 OCD patients seen in the OCD clinic at the National Institute of Mental Health and Neurosciences, Bangalore, were evaluated using the OPCRIT criteria for ICD-10 for the presence of comorbid depressive and anxiety disorders. Results: There were 146 males and 72 females; their mean age at OCD onset was 21.32 ± 0.64 years. Thirty-six (16.5%) patients had depressive episodes, 12 (5.5%) dysthymia and 15 (6.9%) any anxiety disorder. No significant difference in terms of age, sex, marital status or age at onset was found between the OCD patients with and without comorbid anxiety disorder, major depression or dysthymia, except that female OCD patients were more likely than males to have comorbid major depressive disorder. Conclusions: The results of our study are in keeping with previous data from other parts of the world, though the actual rates of comorbidity in our sample appear to be much lower. It remains to be seen whether the differences in rates are a result of methodological issues or different characteristics of sample populations. Further long-term, prospective, methodologically sound studies investigating the comorbidity of depressive and other anxiety disorders in OCD patients are needed to clarify this issue.
Utility of Random Amplification of Polymorphic DNA Assay and BOX-A PCR in Molecular Characterization of Streptococcus pneumoniae Isolates Recovered from Various Ophthalmic Infections
Authors: Kar Upendra K. , Satapathy Gita , Panda S.K. , Das B.K.
Published: October 2005

Background and Objectives: Studies emphasizing the genotype distribution among ophthalmic isolates of Streptococcus pneumoniae are scarce. There is no conclusive information on genetic relatedness of the ophthalmic isolates with the commensal and systemic isolates of S. pneumoniae. Methods: To find out the relatedness between the ophthalmic, invasive and commensal isolates of S. pneumoniae, genotyping was carried out by using RAPD and BOX-A PCR assays. The serotyping was carried out by a rapid co-agglutination method using 12-pooled S. pneumoniae antisera. Results: Topological relationship was identified in order of similarity, and phylogenetic tree was drawn employing sequential clustering alogarithm by UPGMA linkage analysis. The 61 isolates were distributed in 17 RAPD types, 11 BOX-A types and 21 serotypes. Conclusions: Both genotyping techniques were useful in terms of intraspecies genomic variation in S. pneumoniae. BOX-A PCR typing was found to be slightly superior to RAPD analysis in terms of reproducibility and standardization. No specific S. pneumoniae serotype or genotype was found to have special predilection to cause ophthalmic infections.
Beta-Amyloid and Oxidative Stress Jointly Induce Neuronal Death, Amyloid Deposits, Gliosis, and Memory Impairment in the Rat Brain
Authors: Lecanu Laurent , Greeson Janet , Papadopoulos Vassilios
Published: October 2005

Infusion of Fe2+, Aβ42, and buthionine-sulfoximine (FAB), but not Aβ42 alone or in combination with Fe2+, into the left cerebral ventricle of Long-Evans rats for 4 weeks induced memory impairment that was accompanied by increased hyperphosphorylated Tau protein levels in the CSF. FAB-infused animals displayed thioflavin-S-positive amyloid deposits, hyperphosphorylated Tau protein, neuronal loss, and gliosis. Animals treated with Aβ42, Fe2+, or buthionine-sulfoximine alone or in combination failed to show the histological modifications seen with FAB. This data suggests that Aβ42 is not sufficient to induce an Alzheimer’s disease-like symptomatology, and it supports a model whereby a decrease in the brain’s antioxidant defense system leads to the Aβ42-independent oxidative stress necessary for the peptide to induce histopathological changes and memory loss.
Effects of Vitamin E on Phospholipase A2 Activity and Oxidative Damage to the Liver in Streptozotocin-Induced Diabetic Rats
Authors: Rhee Soon-Jae , Jeong Young-Chul , Choi Jeong-Hwa
Published: October 2005

Background: The purpose of the present study was to investigate the effects of vitamin E on oxidative stress and cell membrane fluidity in the liver of streptozotocin (STZ)-induced diabetic rats. Methods: Sprague-Dawley male rats weighing 100 ± 10 g were fed a vitamin E-free diet (the DM-0E group), a 40 mg vitamin E/kg diet (the DM-40E group), or a 400 mg vitamin E/kg diet (the DM-400E group). Result: Dietary vitamin E reduced the increased concentration of lipid peroxides in the liver tissues of diabetic rats through decreasing their increased phospholipase A2 (PLA2) activity and phosphatidylethanolamine hydrolysis. However, vitamin E reduced the accumulation of superoxide radical and decreased the generation of oxidative damage substances, such as the carbonyl value, increased membrane fluidity and lowered oxidative damage. Conclusions: Vitamin E was found to be excellent for regulating the activity of PLA2, reducing the generation of reactive oxygen species and damaging oxidative substances, and maintaining cell membrane fluidity in the liver of diabetic rats.
Long-Term Efficacy and Tolerability of Lanthanum Carbonate: Results from a 3-Year Study
Authors: Hutchison Alastair J. , Maes Bart , Vanwalleghem Johan , Asmus Gernot , Mohamed Elfatih , Schmieder Roland , Backs Wolfgang , Jamar Rene , Vosskühler Andre
Published: October 2005

Background: Control of serum phosphate over the long term is essential in patients with end-stage renal disease. Six-month and 2-year extensions to a 6-month study evaluated the long-term safety, tolerability and efficacy of the new phosphate binder lanthanum carbonate. Methods: Patients who participated in a 6-month, randomized trial comparing lanthanum carbonate with calcium carbonate were eligible for a 24-week, open-label extension. Lanthanum carbonate-treated patients continued taking their established maintenance dose (‘continued-lanthanum group’) and calcium carbonate-treated patients switched to lanthanum carbonate, 375–3,000 mg/day (‘switch group’). Patients could also enter a further 2-year extension. Efficacy parameters, including serum phosphate, were monitored. Results: Mean serum phosphate was ∼1.80 mmol/l throughout the trial. The percentage of patients with controlled serum phosphate (≤1.80 mmol/l) after the 6-month extension was 63.3 and 58.4% in the continued-lanthanum and switch groups, respectively; after the 2-year extension, 54.4% of patients had controlled serum phosphate. After discontinuation of calcium carbonate and initiation of lanthanum carbonate, the hypercalcemia incidence was 2.7%, compared with 20.2% during the double-blind phase. Calcium × phosphate product was maintained at an acceptable level. Lanthanum carbonate was well tolerated; adverse events were mild/moderate and mainly gastrointestinal. Conclusions: Lanthanum carbonate maintains effectiveness with continued tolerability for up to 3 years.
Ultrasonic Morphological Characteristics of Yolk Sac in Pregnancy Complicated with Type-1 Diabetes mellitus
Authors: Ivanišević Marina , Djelmiš Josip , Jalsovec Dubravko , Bljajic Danko
Published: October 2005

Background/Aims: In pregnancy complicated with type-1 diabetes mellitus (DM), information on the predictive value of yolk sac (YS) measurement and YS morphology on outcome has been scant. The aim of this study was to compare the size and shape of the YS using transvaginal sonography (TVS) in normal pregnancies and those complicated with well-regulated type-1 DM. Methods: Sixty healthy pregnant women and 60 pregnant women with type-1 DM underwent TVS. The correlation coefficients between YS and gestational age (GA), YS and crown-rump length (CRL), and YS and human chorionic gonadotropin (HCG) were calculated. Statistical analysis showed that the difference between the YS diameters for each gestational week among the groups studied was highly statistically significant. Statistical analysis of the results was performed with SPSS 10 software. Results: A highly statistically positive correlation was found between YS diameter and GA, CRL and HCG levels for both groups studied. The YS diameter in type-1 DM pregnancies was statistically significantly larger for GA than that of healthy pregnant women. The YS diameter in type-1 DM women was statistically significantly larger after 6 weeks of gestation and the HbA1c levels were >6% when compared to the YS dimensions in the group with HbA1c concentrations of <6%. Conclusion:A gross change in YS size may indicate or reflect significant dysfunction of the maternofetal transport system in early pregnancy which is complicated with type-1 DM.
Investigation of β-Lactamases in Clinical Isolates of Staphylococcus aureus for Further Explanation of Borderline Methicillin Resistance
Authors: Keserű Judit Szilvia , Gál Zsuzsanna , Barabás György , Benkő Ilona , Szabó István
Published: October 2005

Background: Borderline methicillin resistance in Staphylococcus aureus is due to β-lactamase overproduction and/or specific methicillinases. Methods: β-Lactamase activity in culture supernatants and in cytoplasmic membrane fractions was estimated by bioassay and by SDS-PAGE combined with nitrocefin assay. Results: During the investigation of borderline methicillin-resistant Staphylococcus aureus (BORSA) strains VU94 and 822 two β-lactamases were detected in the membranes, with molecular weights of 13 and 30 kDa. The latter could be found in the culture supernatants, too. In the presence of globomycin, this enzyme disappeared from the membrane, and the oxacillin-hydrolyzing activity of the membrane decreased to the level of susceptible strains. Both β-lactamases were detected in the methicillin-resistant Staphylococcus aureus strain studied, but the susceptible strains possessed only the first enzyme. Conclusions: The 30-kDa β-lactamase proved to be a methicillinase, and it can be one of the main causes of the borderline phenotype of BORSA strains. The other enzyme is one of the smallest β-lactamases published to date.
Adjuvant Systemic Chemotherapy with Gemcitabine for Stage IV Pancreatic Cancer: A Preliminary Report of Initial Experience
Authors: Kurosaki Isao , Hatakeyama Katsuyoshi
Published: October 2005

Background: The effects of gemcitabine on postoperative adjuvant chemotherapy were evaluated in patients with stage IV pancreatic cancer. The results were compared with those of our historical control patients treated with surgery alone. Patients and Methods: Nineteen patients with stage IV pancreatic cancer who had pancreatic resection with curative intent during the 5 years up to February 2003 were enrolled in this study. Nine cases received postoperative adjuvant chemotherapy with biweekly administration of 1,000 mg/m2 gemcitabine, while the remaining 10 cases underwent surgery without any adjuvant chemotherapy. Results: The chemotherapy was well tolerated with only mild symptomatic and hematologic toxicities. The disease-specific cumulative survival rates of the chemotherapy and surgery-alone groups were 86 and 70% at 1 year, and 50 and 12% at 2 years, with a median survival of 20 and 14 months, respectively (p = 0.0255). The disease-free interval was improved, and the occurrence of hepatic metastasis was reduced in the chemotherapy group compared with the surgery-alone group. Conclusions: Adjuvant systemic chemotherapy utilizing gemcitabine was feasible with acceptable adverse effects, and showed some survival benefit in stage IV pancreatic cancer patients. Further investigation into gemcitabine-based combination therapies is warranted.
Intrapulmonary Concentrations of Telithromycin: Clinical Implications for Respiratory Tract Infections due to Streptococcus pneumoniae
Authors: Ong C.T. , Dandekar P.K. , Sutherland C. , Nightingale C.H. , Nicolau D.P.
Published: October 2005

Background: Antimicrobial efficacy is dependent on the ability of the agent to reach the site of infection. To assess the bronchopulmonary drug disposition of a novel ketolide, telithromycin (TEL), the epithelial lining fluid (ELF) and alveolar macrophage (AM) concentrations were utilized as a surrogate marker for lung penetration. Methods: Adult subjects scheduled for diagnostic bronchoscopy received oral TEL 800 mg once daily for 5 days. Plasma and bronchoalveolar lavage (BAL) samples were collected 2, 8, 12, or 24 h after the last TEL dose. TEL concentrations in the ELF and AM were determined using a validated HPLC assay. ELF drug concentrations were calculated using the urea dilution method. Results: Seventeen subjects with a mean age 65 ± 13 years and a mean weight of 81 ± 25 kg completed this open-label study. The median (range) TEL concentrations in plasma and ELF, respectively, were 1.09 mg/l (1.00–4.81) and 3.91 mg/l (2.64–9.59) at 2 h (n = 6), 0.48 and 1.09 mg/l at 8 h (n = 1), 0.65 mg/l (0.18–1.55) and 1.81 mg/l (0.61–10.0) at 12 h (n = 5), and 0.11 mg/l (0.09–0.24) and 0.69 mg/l (0.15–1.58) at 24 h (n = 5). The median AM concentrations obtained from these subjects were 53.35 mg/l at 2 h, 32.55 mg/l at 8 h, 65.96 mg/l at 12 h, and 26.43 mg/l at 24 h. Overall TEL was well tolerated. No discontinuation was required due to an adverse event. Conclusions: TEL displayed high intrapulmonary penetration with ELF concentrations exceeding that of plasma at all time points. AM intracellular concentrations were multiple times higher than in the ELF and plasma. These data support the clinical efficacy of TEL against intracellular and extracellular pathogens, particularly with Streptococcus pneumoniae having an MIC90 well below achievable concentrations at the site of infection.
Changes in Ubiquitin Proteasome Pathway Gene Expression in Skeletal Muscle With Exercise and Statins
Authors: Urso Maria L. , Clarkson Priscilla M. , Hittel Dustin , Hoffman Eric P. , Thompson Paul D.
Published: October 2005

Objective—

Statins are safe medications but have side effects including myalgia and rhabdomyolysis. How statins provoke muscle damage is not known, but this effect is exacerbated by exercise.

Methods and Results—

Healthy subjects took Atorvastatin (80 mg/daily) or placebo for 4 weeks. Biopsies of both vastus lateralis muscles were performed 8 hours after eccentric exercise (known to result in muscle soreness and damage) of the left leg at baseline and the right leg after statin/placebo treatment. Gene expression was determined using Affymetrix GeneChips, and selected genes confirmed by polymerase chain reaction (qRT-PCR). Atorvastatin had little effect on gene expression at rest. When combined with exercise, 56 genes were differentially expressed with 18% involved in the ubiquitin proteasome pathway (UPP) and 20% involved in protein folding and catabolism, and apoptosis.

Conclusion—

This is the first investigation to our knowledge to implicate involvement of the UPP in skeletal muscle in response to combined exercise and statin treatment, possibly explaining the onset of myalgia with exertion. Statins may alter the response of muscle to exercise stress by altering the action of the UPP, protein folding, and catabolism, disrupting the balance between protein degradation and repair.
Therapeutic Angiogenesis With Intramuscular Injection of Low-Dose Recombinant Granulocyte-Colony Stimulating Factor
Authors: Lee Mejeong , Aoki Mika , Kondo Takahisa , Kobayashi Koichi , Okumura Kenji , Komori Kimihiro , Murohara Toyoaki
Published: October 2005

Objective—

In vivo administration of granulocyte colony-stimulating factor (G-CSF) has been shown to facilitate regeneration of cardiovascular tissues. However, G-CSF causes marked leukocytosis that potentially induces adverse cardiovascular events. Earlier studies showed that G-CSF had direct stimulatory actions on mature endothelial cells, resulting in promotion of angiogenesis. We thus examined whether low doses of recombinant human G-CSF (rhG-CSF) locally injected into ischemic tissues would stimulate angiogenesis without inducing severe leukocytosis.

Methods and Results—

Reverse-transcription polymerase chain reaction (PCR) revealed expression of G-CSF receptor in human umbilical vein endothelial cells (HUVECs). rhG-CSF (100 ng/mL) enhanced migration and tube formation but not proliferation of HUVECs in vitro. We then examined the effects of rhG-CSF on angiogenesis in a rat model of hindlimb ischemia. Nude rats received in their ischemic skeletal muscles either rhG-CSF (2, 10, 20 μg/kg per day) or saline (control) for 6 days. Laser Doppler blood flowmetry (LDBF) revealed an augmented ischemic/normal limb LDBF ratio and an increased capillary density in the rhG-CSF–treated groups compared with the control at days 14, 21, and 28 (
P
<0.05). These doses of rhG-CSF induced only mild leukocytosis (≈1.4-fold increases versus baseline).

Conclusions—

rhG-CSF promoted endothelial migration and tube formation in vitro. Local injection of low doses rhG-CSF effectively augmented ischemia-induced angiogenesis in vivo. This treatment regimen of low-dose rhG-CSF may become a new and safe modality for therapeutic angiogenesis.
Tumor Necrosis Factor-α–Induced Iron Sequestration and Oxidative Stress in Human Endothelial Cells
Authors: Nanami Masayoshi , Ookawara Tomomi , Otaki Yoshinaga , Ito Katsukiyo , Moriguchi Rintarou , Miyagawa Koji , Hasuike Yukiko , Izumi Masaaki , Eguchi Hironobu , Suzuki Keiichiro , Nakanishi Takeshi
Published: October 2005

Objective—

Tumor necrosis factor (TNF)-α–induced endothelial injury, which is associated with atherosclerosis, is mediated by intracellular reactive oxygen species. Iron is essential for the amplification of oxidative stress. We tested whether TNF-α accelerated iron accumulation in vascular endothelium, favoring synthesis of hydroxyl radical.

Methods and Results—

Diverse iron transporters, including iron import proteins (transferrin receptor [TfR] and divalent metal transporter 1 [DMT1]) and an iron export protein (ferroportin 1 [FP1]) coexist in human umbilical endothelial cells (HUVECs). TNF-α caused upregulation of TfR and DMT1 and downregulation of FP1, which were demonstrated in mRNA as well as protein levels. These changes in iron transporters were accompanied by accumulation of iron that was both transferrin-dependent and transferrin-independent. Modifications of these mRNAs were regulated post-transcriptionally, and were coordinated with activation of binding activity of iron regulatory protein 1 to the iron responsive element on transporter mRNAs. Using a salicylate trap method, we observed that only simultaneous exposure of endothelial cells to iron and TNF-α accelerated hydroxyl radical production.

Conclusions—

TNF-α could cause intracellular iron sequestration, which may participate importantly in the pathophysiology of atherosclerosis and cardiovascular disease.
Effect of Fenofibrate on Plasma Lipoprotein Composition and Kinetics in Patients With Complete Hepatic Lipase Deficiency
Authors: Ruel Isabelle L. , Lamarche Benoît , Mauger Jean-François , Badellino Karen O. , Cohn Jeffrey S. , Marcil Michel , Couture Patrick
Published: October 2005

Objective—The goal of this study was to characterize the effect of microcoated fenofibrate (160 mg/day for 6 months) on plasma lipoprotein composition and kinetics in 2 patients with complete hepatic lipase (HL) deficiency.

Methods and Results—Fenofibrate treatment normalized the plasma lipoprotein profile of patients with complete HL deficiency, as evidenced by significant reductions in the plasma concentration of cholesterol (−49%) and triglycerides (−82%) and a significant increase in low-density lipoprotein (LDL) size (251.5±1.8 versus 263.5±0.7 Å). The in vivo kinetics of very low–density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), and LDL apolipoprotein (apo)B-100 and plasma apoA-I and apoA-II were studied using a primed-constant infusion of L-[5,5,5-D3]-leucine for 12 hours in the fasted state. Fenofibrate treatment in complete HL-deficient patients substantially decreased plasma concentrations of VLDL, IDL, and LDL apoB-100 attributable to important increases in VLDL (+325%), IDL (+129%), and LDL (+218%) apoB-100 fractional catabolic rates (FCR). IDL apoB-100 FCR nevertheless remained 60% lower after treatment compared with values obtained in controls (n=5). The kinetics of plasma apoA-I and apoA-II as well as the capacity of total plasma and of high-density lipoprotein particles to efflux cellular cholesterol from normal human skin fibroblasts was not altered by fenofibrate.

Conclusion—Fenofibrate therapy exerts a pronounced antiatherogenic effect on triglyceride-rich lipoproteins even in the complete absence of HL.
Novel Autologous Cell Therapy in Ischemic Limb Disease Through Growth Factor Secretion by Cultured Adipose Tissue–Derived Stromal Cells
Authors: Nakagami Hironori , Maeda Kazuhisa , Morishita Ryuichi , Iguchi Sota , Nishikawa Tomoyuki , Takami Yoichi , Kikuchi Yasushi , Saito Yukihiro , Tamai Katsuto , Ogihara Toshio , Kaneda Yasufumi
Published: October 2005

Objective—

The delivery of autologous progenitor cells into ischemic tissue of patients is emerging as a novel therapeutic option. Here, we report the potential impact of cultured adipose tissue–derived cells (ADSC) on angiogenic cell therapy.

Method and Results—

ADSC were isolated from C57Bl/6 mouse inguinal adipose tissue and showed high expression of
Sca
I and CD44, but not c-kit, Lin, CD34, CD45, CD11b, and CD31, compatible with that of mesenchymal stem cells from bone marrow. In coculture conditions with ADSC and human aortic endothelial cells (ECs) under treatment with growth factors, ADSC significantly increased EC viability, migration and tube formation mainly through secretion of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). At 4 weeks after transplantation of ADSC into the ischemic mouse hindlimb, the angiogenic scores were improved in the ADSC-treated group, which were evaluated with blood flow by laser Doppler imaging (LDI) and capillary density by immunostaining with anti-CD31 antibody. However, injected ADSC did not correspond to CD31, von Willebrand factor, and α-smooth muscle actin-positive cells in ischemic tissue.

Conclusion—

These adipose tissue–derived cells demonstrated potential as angiogenic cell therapy for ischemic disease, which appears to be mainly achieved by their ability to secrete angiogenic growth factors.
Myocardin Enhances Smad3-Mediated Transforming Growth Factor-β
1
Signaling in a CArG Box-Independent Manner
Authors: Qiu Ping , Ritchie Raquel P. , Fu Zhiyao , Cao Dongsun , Cumming Jerry , Miano Joseph M. , Wang Da-Zhi , Li Hui J. , Li Li
Published: October 2005

Transforming growth factor (TGF)-β
1
is an important cytokine involved in various diseases. However, the molecular mechanism whereby TGF-β
1
signaling modulates the regulatory network for smooth muscle gene transcription remains largely unknown. To address this question, we previously identified a Smad-binding element (SBE) in the SM22α promoter as one of the TGF-β
1
response elements. Here, we show that mutation of the SBE reduces the activation potential of a SM22α promoter in transgenic mice during embryogenesis. Chromatin immunoprecipitation assays reveal that TGF-β
1
induces Smad3 binding to the SM22α promoter in vivo. A multimerized SBE promoter responsive to TGF-β
1
signaling is highly activated by Smad3 but not by the closely related Smad2. Intriguingly, myocardin (Myocd), a known CArG box-dependent serum response factor coactivator, participates in Smad3-mediated TGF-β
1
signaling and synergistically stimulates Smad3-induced SBE promoter activity independent of the CArG box; no such synergy is seen with Smad2. Importantly, Myocd cooperates with Smad3 to activate the wild-type SM22α, SM myosin heavy chain, and SMα-actin promoters; they also activate the CArG box-mutated SM22α promoter as well as the CArG box-independent aortic carboxypeptidase-like protein promoter. Immunopreciptiation assays reveal that Myocd and Smad3 directly interact both in vitro and in vivo. Mutagenesis studies indicate that the C-terminal transactivation domains of Myocd and Smad3 are required for their functional synergy. These results reveal a novel regulatory mechanism whereby Myocd participates in TGF-β
1
signal pathway through direct interaction with Smad3, which binds to the SBEs. This is the first demonstration that Myocd can act as a transcriptional coactivator of the smooth muscle regulatory network in a CArG box-independent manner.
Bone Morphogenetic Protein Signaling Modulates Myocardin Transactivation of Cardiac Genes
Authors: Callis Thomas E. , Cao Dongsun , Wang Da-Zhi
Published: October 2005

Bone morphogenetic proteins (BMPs) play important roles in cardiovascular development. However, how BMP-signaling pathways regulate cardiac gene expression is less clear. We have previously identified myocardin as a cardiac and smooth muscle-specific transcriptional cofactor for serum response factor (SRF). Myocardin potently activates target gene expression by tethering with SRF bound to SRF-responsive elements, the CArG box. Here, we show that Smad1, an effector of the BMP-signaling pathway, synergistically activates myocardin-dependent cardiac gene expression. Interestingly, the CArG box is necessary and sufficient to mediate such synergy, whereas no obvious Smad-binding element appears to be involved. Consistent with their functional interaction, we find that myocardin and Smad1 proteins interact directly. Furthermore, myocardin protein levels were dramatically increased by BMP-2 treatment in cardiomyocytes. These findings suggest myocardin participates in a BMP signaling-dependent cardiac gene transcriptional program.
Cerebral Blood Flow Thresholds for mRNA Synthesis After Focal Ischemia and the Effect of MK-801
Authors: Kamiya Tatsushi , Jacewicz Michael , Nowak Thaddeus S. , Pulsinelli William A.
Published: October 2005

Background and Purpose—

MK-801 is a noncompetitive antagonist of N-methyl-
d
-aspartate subtype glutamate receptors with protective efficacy in experimental stroke. This study examined the impact of MK-801 on cerebral blood flow (CBF) and its relationship to gene expression changes during focal ischemia.

Methods—

Spontaneously hypertensive rats were subjected to surgical occlusion of the middle cerebral artery and ipsilateral common carotid artery after 30 minutes pretreatment with 5 mg/kg MK-801 or saline vehicle. After 2.5 hours of ischemia, regional CBF was evaluated by [
14
C]iodoantipyrine autoradiography and compared with distributions of gene expression changes evaluated by in situ hybridization detection of mRNAs encoding several immediate–early genes and the stress protein, hsp72.

Results—

MK-801 increased CBF in contralateral cortex from 93±15 to 187±37 mL/100 g per minute and produced a significant 25% reduction in the volume of ischemic cortex ipsilateral to occlusion. The extent of cortex failing to express inducible mRNAs correspondingly decreased, but the CBF threshold for mRNA synthesis remained unchanged (25 to 30 mL/100 g per minute). Widespread immediate–early gene expression in the neocortex became restricted to periinfarct regions after MK-801 treatment, and hybridization patterns in the striatum and hippocampus reflected the altered topography of cortical activation after drug treatment.

Conclusions—

MK-801 alters ischemia-induced gene expression by 2 distinct mechanisms. Generalized increases in CBF reduce the volume of cortex falling below ischemic injury thresholds, protecting tissue and facilitating transcription of inducible genes proximal to the ischemic focus. In addition, MK-801 attenuates the signals that induce expression of immediate–early genes in cortical and subcortical regions remote from the middle cerebral artery territory.
Shape and Volume of Lacunar Infarcts
Authors: Hervé Dominique , Mangin Jean-François , Molko Nicolas , Bousser Marie-Germaine , Chabriat Hugues
Published: October 2005

Background and Purpose—

The shape and exact size of lacunar infarcts have been investigated only postmortem. Recent imaging techniques based on triangulation and connectivity can now be used for 3D segmentation of cerebral lesions. The shape and size of lacunar infarcts was investigated using these techniques in 10 cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) patients.

Methods—

We segmented 102 lacunar infarcts on T1-weighted images. The surface of the corresponding set of voxels was computed as a mesh of triangles. Thereafter, the shape of each lesion in 3D was visually analyzed by 2 investigators.

Results—

The volume of lesions ranged from 10.5 to 1146 mm, with 93% of them having a volume <500 mm; 83% lacunar infarcts had a spheroid or ovoid shape, but 17% presented as sticks, slabs, or with a complex shape. Lesions with multiple components appeared larger than the others, and a tail extension was noticed in 13 of 102 lesions.

Conclusions—

These results suggest the following: (1) most lacunar infarcts in CADASIL have a volume far below one third of that of a sphere of 15 mm in diameter, the upper limit currently used for their identification on 2D imaging; (2) a significant proportion of lacunar infarcts have a shape distinct from the spheroid-ovoid morphology; and (3) lesions with a complex shape may result from the involvement of the largest small arteries, confluence of ischemic lesions, or secondary tissue degeneration. The segmentation of lacunar infarcts appears promising to better understand the pathophysiology of tissue lesions secondary to small vessel diseases.
Variation in the Carotid Bifurcation Geometry of Young Versus Older Adults
Authors: Thomas Jonathan B. , Antiga Luca , Che Susan L. , Milner Jaques S. , Hangan Steinman Dolores A. , Spence J. David , Rutt Brian K. , Steinman David A.
Published: October 2005

Background and Purpose—

Retrospective analysis of clinical data has demonstrated major variations in carotid bifurcation geometry, in support of the notion that an individual’s vascular anatomy or local hemodynamics may influence the development of atherosclerosis. On the other hand, anecdotal evidence suggests that vessel geometry is more homogenous in youth, which would tend to undermine this geometric risk hypothesis. The purpose of our study was to test whether the latter is indeed the case.

Methods—

Cross-sectional images of the carotid bifurcations of 25 young adults (24±4 years) and a control group of 25 older subjects (63±10 years) were acquired via MRI. Robust and objective techniques were developed to automatically characterize the 3D geometry of the bifurcation and the relative dimensions of the internal, external, and common carotid arteries (ICA, ECA, and CCA, respectively).

Results—

Young vessels exhibited significantly less interindividual variation in the following geometric parameters: bifurcation angle (48.5±6.3° versus 63.6±15.4°); ICA angle (21.6±6.7° versus 29.2±11.3°); CCA tortuosity (0.010±0.003 versus 0.014±0.011); ICA tortuosity (0.025±0.013 versus 0.086±0.105); ECA/CCA diameter ratio (0.81±0.06 versus 0.75±0.13), ICA/CCA (0.81±0.06 versus 0.77±0.12) diameter ratio, and bifurcation area ratio (1.32±0.15 versus 1.19±0.35).

Conclusions—

The finding of more modest interindividual variations in young adults suggests that, if there is a geometric risk for atherosclerosis, its early detection may prove challenging. Taken together with the major interindividual variations seen in older vessels, it suggests a more complex interrelationship between vascular geometry, local hemodynamics, vascular aging, and atherosclerosis, the elucidation of which now calls for prospective studies.
Economic Benefit of Increasing Utilization of Intravenous Tissue Plasminogen Activator for Acute Ischemic Stroke in the United States
Authors: Demaerschalk Bart M. , Yip Todd R.
Published: October 2005

Background and Purpose—

Health economic analyses of intravenous tissue plasminogen activator (tPA) in acute ischemic stroke reveal a substantial cost savings. Unfortunately, tPA is vastly underused. The purpose of this study was to determine the economic impact of increasing tPA utilization in the United States.

Methods—

Annual incidence estimates of ischemic stroke in the United States and individual states were obtained. The proportion of all ischemic stroke patients who receive tPA was derived from published data. Economic analyses that report the expected annual cost savings of tPA were consulted. The analysis was conducted from the perspective of the healthcare system over a time period of 1 year. With incremental increases in the proportion of all ischemic stroke patients treated with tPA, potential cost savings were recalculated. The outcomes are expressed in dollars saved annually.

Results—

There are 616 000 new ischemic stroke patients annually. A $600 net cost savings is associated with each tPA-treated patient. Currently, an estimated 2% of all ischemic stroke patients receive tPA. If the proportion was increased to 4, 6, 8, 10, 15, or 20%, the realized cost savings would be approximately $15, 22, 30, 37, 55, and 74 million, respectively.

Conclusions—

If even small manageable increases in the proportion of all ischemic stroke patients who received tPA were achieved, it would result in an enormous realized savings for America’s healthcare system.
Determinants of Staged Endovascular and Surgical Treatment Outcome of Brain Arteriovenous Malformations
Authors: Hartmann Andreas , Mast Henning , Mohr Jay P. , Pile-Spellman John , Connolly E. Sander , Sciacca Robert R. , Khaw Alexander , Stapf Christian
Published: October 2005

Background and Purpose—

Therapy of brain arteriovenous malformations (AVMs) often requires the combination of different treatment modalities. Independently assessed data on neurologic outcome after multidisciplinary AVM therapy are scarce.

Methods—

The 119 consecutive patients (49% women, mean age 34±13 years) with brain AVMs receiving endovascular embolization followed by surgical treatment were analyzed. Neurologic impairment was assessed prospectively by a neurologist using the modified Rankin Scale (mRS) before, during, and after completed AVM therapy. The association of demographic, clinical, and morphologic characteristics with new treatment-related neurologic deficits was calculated.

Results—

The 119 patients were treated with 240 superselective embolizations (median, 2; range, 1 to 8) using n-butyl cyanoacrylate. Mean follow-up time after surgery was 9.6±13.2 months. On the Spetzler-Martin scale, 8% of the AVMs were grade 1, 27% grade 2, 40% grade 3, 22% grade 4, and 3% grade 5. Disabling treatment-related complications (mRS≥3) occurred in 5% (95% confidence interval [CI], 1% to 9%) of the patients. Nondisabling new deficits were observed in another 42% (95% CI, 33% to 51%). No patient died. Nonhemorrhagic AVM presentation (odds ratio [OR], 5.00; 95% CI, 1.75 to 14.29), deep venous drainage (OR, 3.09; 95% CI, 1.43 to 6.64), AVM location in an eloquent brain region (OR, 2.42; 95% CI, 1.10 to 5.33), and large AVM size (OR, 1.05; 95% CI, 1.01 to 1.09) were independently associated with new treatment-related deficits.

Conclusions—

Our results suggest an increased treatment risk for patients with previously unbled AVMs from combined endovascular and surgical AVM therapy. Additional risk factors for treatment-related neurologic deficits may be large AVM size, deep venous drainage, and AVM location in eloquent brain regions.
NIHSS Training and Certification Using a New Digital Video Disk Is Reliable
Authors: Lyden Patrick , Raman Rema , Liu Lin , Grotta James , Broderick Joseph , Olson Scott , Shaw Sandi , Spilker Judith , Meyer Brett , Emr Marian , Warren Margo , Marler John
Published: October 2005

Background and Purpose—

NIH Stroke Scale certification is required for participation in modern stroke clinical trials and as part of good clinical care in stroke centers. The existing training and certification videotapes, however, are more than 10 years old and do not contain an adequate balance of patient findings.

Methods—

After producing a new NIHSS training and demonstration DVD, we selected 18 patients representing all possible scores on 15 scale items for a new certification DVD. Patients were divided into 3 certification groups of 6 patients each, balanced for lesion side, distribution of scale item findings, and total score. We sought to measure interrater reliability of the certification DVD using methodology previously published for the original videotapes. Raters were recruited from 3 experienced stroke centers. Each rater watched the new training DVD and then evaluated one of the 3 certification groups.

Results—

Responses were received from 112 raters: 26.2% of all responses came from stroke nurses, 34.1% from emergency departments/other physicians, and 39.6% from neurologists. One half (50%) of raters were previously NIHSS-certified. Item responses were tabulated, scoring performed as previously published, and agreement measured with unweighted κ coefficients for individual items and an intraclass correlation coefficient for the overall score. κ ranged from 0.21±0.05 (ataxia) to 0.92±0.09 (LOC-C questions). Of 15 items, 2 showed poor, 11 moderate, and 2 excellent agreement based on κ scores. The intraclass correlation coefficient for total score was 0.94 (95% confidence interval, 0.84 to 1.00). Reliability scores were similar among specialists and centers, and there were no differences between nurses and physicians. κ scores trended higher among raters previously certified.

Conclusions—

These certification DVDs are reliable for NIHSS certification, and scoring sheets have been posted on a web site for real-time, online certification.
Absence of Microemboli on Transcranial Doppler Identifies Low-Risk Patients With Asymptomatic Carotid Stenosis
Authors: Spence J. David , Tamayo Arturo , Lownie Stephen P. , Ng Wai P. , Ferguson Gary G.
Published: October 2005

Background and Purpose—

Carotid endarterectomy clearly benefits patients with symptomatic severe stenosis (SCS), but the risk of stroke is so low for asymptomatic patients (ACS) that the number needed to treat is very high. We studied transcranial Doppler (TCD) embolus detection as a method for identifying patients at higher risk who would have a lower number needed to treat.

Methods—

Patients with carotid stenosis of ≥60% by Doppler ultrasound who had never been symptomatic (81%) or had been asymptomatic for at least 18 months (19%) were studied with TCD embolus detection for up to 1 hour on 2 occasions a week apart; patients were followed for 2 years.

Results—

319 patients were studied, age (standard deviation) 69.68 (9.12) years; 32 (10%) had microemboli at baseline (TCD+). Events were more likely to occur in the first year. Patients with microemboli were much more likely to have microemboli 1 year later (34.4 versus 1.4%;
P
<0.0001) and were more likely to have a stroke during the first year of follow-up (15.6%, 95% CI, 4.1 to 79; versus 1%, 95% CI, 1.01 to 1.36;
P
<0.0001).

Conclusions—

Our findings indicate that TCD− ACS will not benefit from endarterectomy or stenting unless it can be done with a risk <1%; TCD+ may benefit as much as SCS if their surgical risk is not higher. These findings suggest that ACS should be managed medically with delay of surgery or stenting until the occurrence of symptoms or emboli.