bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement in Blood Pressure With Inhibition of the Epithelial Sodium Channel in Blacks With Hypertension
Authors: Saha Chandan , Eckert George J. , Ambrosius Walter T. , Chun Tae-Yon , Wagner Mary Anne , Zhao Qianqian , Pratt J. Howard
Published: August 2005

Hypertension in blacks is more prevalent and less often controlled than the hypertension of other ethnic groups. We sought to explore the benefit of adding inhibitors of the epithelial sodium channel (ENaC), an aldosterone-regulated site of sodium reabsorption in the distal nephron, to the antihypertensive regimen of black hypertensive patients. In a prospective, randomized, placebo-controlled, double-blind clinical trial, we used a 2-by-2 factorial design with 4 treatment groups: amiloride (a direct inhibitor of ENaC), spironolactone (an aldosterone receptor antagonist), the combination of both drugs, and placebo. The subjects (n=98) had an elevated blood pressure despite treatment that included a diuretic and a calcium channel blocker; the level of plasma renin activity was ≤0.56 ng/L per second. The primary end points were changes from baseline in systolic and diastolic blood pressure over a 9-week period of treatment. The reductions in systolic and diastolic blood pressures (mm Hg) were, respectively, 9.8±1.6 (SE) and 3.4±1.0 for amiloride (
P
<0.001) and 4.6±1.6 (
P
=0.006) and 1.8±1.0 for spironolactone (
P
=0.07). Treatment with either amiloride or spironolactone or the combination was well tolerated; no patient experienced hyperkalemia. In a substudy, plasma endothelin-1 levels were observed to decrease after 3 weeks of treatment with spironolactone (
P
<0.001), consistent with a non–ENaC-related potential benefit of spironolactone. In conclusion, treatment with either amiloride or spironolactone can provide an additional reduction in blood pressure in blacks already receiving conventional antihypertensive therapy.
C-Reactive Protein as a Predictor of Cardiovascular Risk in a Population With a High Prevalence of Diabetes
Authors: Best Lyle G. , Zhang Ying , Lee Elisa T. , Yeh Jeun-Liang , Cowan Linda , Palmieri Vittorio , Roman Mary , Devereux Richard B. , Fabsitz Richard R. , Tracy Russell P. , Robbins David , Davidson Michael , Ahmed Aftab , Howard Barbara V.
Published: August 2005

Background—

High-sensitivity C-reactive protein (CRP) has been investigated extensively as a marker for predicting the risk of cardiovascular disease (CVD). CVD accounts for a large proportion of mortality and morbidity in American Indians; we sought to test the association of CRP and CVD in a population-based American Indian cohort 45 to 74 years old.

Methods and Results—

Of 3277 participants who were CVD-free at baseline, 542 had CRP >10 mg/L and were excluded from analysis; 50.1% of those included had diabetes. There were 343 CVD events among this cohort during a median follow-up of 6.2 years. Multiple CVD risk factors were used as covariates in Cox proportional-hazard models. After exclusions, the median CRP (3.2 mg/L) was higher than reported in many other populations. CRP predicted CVD in models adjusted for traditional risk factors, but not when albuminuria and fibrinogen were included. In subgroup analysis, CRP was strongly related to incident CVD among nondiabetic women participants, even after adjustment for traditional CVD risk factors and other indicators of inflammation. Conversely, CRP was elevated beyond the useful range of the American Heart Association/Centers for Disease Control and Prevention clinical guidelines in 16% of this population, and CRP was not predictive of CVD in important subgroups, such as those with diabetes.

Conclusions—

CRP was a predictor of CVD in this American Indian population with a high prevalence of diabetes and other risk factors. The predictive ability of CRP varies considerably among subgroups with different risk factor profiles.
Inactivation of Monocarboxylate Transporter MCT3 by DNA Methylation in Atherosclerosis
Authors: Zhu Shoukang , Goldschmidt-Clermont Pascal J. , Dong Chunming
Published: August 2005

Background—

Monocarboxylate transporters (MCTs) mediate lactate transport across the plasma membrane of cells. The molecular mechanisms regulating monocarboxylate transport in smooth muscle cells (SMCs) remain poorly characterized. The aim of this study was to investigate the effects of DNA methylation on MCT expression and lactate transport in SMCs in relation to atherosclerosis.

Methods and Results—

MCT expression was determined by real-time reverse transcription–polymerase chain reaction, Western blotting, and immunohistochemistry in SMCs isolated from human aortas and coronary arteries. Bisulfite sequencing and confocal microscopic analysis were used to study DNA methylation and lactate transport in SMCs, respectively. Downregulation of MCT3 and impaired lactate transport were detected in proliferating/synthetic SMCs, relative to the contractile phenotype. A passage number– and atherosclerotic lesion–dependent methylation pattern of MCT3 was demonstrated in the CpG island located in exon 2. Treatment of SMCs with the demethylating agent 5-aza-2′-deoxycytidine restored MCT3 expression and normalized lactate transport. Furthermore, small interfering RNA–mediated specific MCT3 knockdown substantially stimulated SMC proliferation.

Conclusions—

These data indicate that DNA methylation may modify monocarboxylate transport by suppressing MCT3 expression, which could be important in regulating SMC function and the development of atherosclerosis.
Impact of a Single Intravenous Administration of Nicorandil Before Reperfusion in Patients With ST-Segment–Elevation Myocardial Infarction
Authors: Ishii Hideki , Ichimiya Satoshi , Kanashiro Masaaki , Amano Tetsuya , Imai Kenji , Murohara Toyoaki , Matsubara Tatsuaki
Published: August 2005

Background—

Intravenous nicorandil, a hybrid compound of ATP-sensitive potassium channel opener and nitric oxide donor, has been reported to ameliorate early functional and clinical problems in patients with acute myocardial infarction. However, its effects on the late phase remain unclear.

Methods and Results—

This follow-up study to 5 years of a randomized, double-blinded trial was conducted among 368 patients with first ST-segment–elevation myocardial infarction undergoing percutaneous coronary intervention (PCI). They were randomly assigned to receive 12 mg of nicorandil or a placebo intravenously just before reperfusion. We analyzed incidence of cardiovascular death or rehospitalization for congestive heart failure after PCI as well as various aspects of epicardial flow and microvascular function. Mean follow-up was 2.4 years (SD, 1.4). A total of 12 (6.5%) patients receiving nicorandil and 30 (16.4%) receiving placebo had cardiovascular death or hospital admission for congestive heart failure (hazard ratio, 0.39; 95% CI, 0.20 to 0.76;
P
=0.0058). Postprocedural TIMI 3 flow was obtained in 89.7% of the nicorandil group and in 81.4% of the placebo (hazard ratio, 1.99; 95% CI, 1.09 to 3.65;
P
=0.025). Corrected TIMI frame count was furthermore lower in the nicorandil group (21.0±9.1 versus 25.1±14.1;
P
=0.0009). ST-segment resolution >50% was observed in 79.5% and 61.2% of the nicorandil and placebo groups, respectively (hazard ratio, 2.45; 95% CI, 1.54 to 3.90;
P
=0.0002).

Conclusions—

The addition of intravenous nicorandil to PCI leads to beneficial clinical outcomes and prevents cardiovascular events of long duration and death in patients with ST-segment–elevation myocardial infarction.
Impact of the Pravastatin or Atorvastatin Evaluation and Infection Therapy–Thrombolysis In Myocardial Infarction 22/Reversal of Atherosclerosis With Aggressive Lipid Lowering Trials on Trends in Intensive Versus Moderate Statin Therapy in Ontario, Canada
Authors: Austin Peter C. , Mamdani Muhammad M.
Published: August 2005

Background—

In March 2004, the Reversal of Atherosclerosis With Aggressive Lipid Lowering (REVERSAL) trial demonstrated that intensive lipid-lowering therapy (atorvastatin 80 mg/d) reduced progression of coronary atherosclerosis compared with moderate lipid-lowering therapy (pravastatin 40 mg/d). The following month, the Pravastatin or Atorvastatin Evaluation and Infection Therapy–Thrombolysis In Myocardial Infarction 22 (PROVE IT–TIMI 22) study demonstrated the superiority of intensive (atorvastatin 80 mg/d) versus moderate (pravastatin 40 mg/d) lipid-lowering therapy for reducing death or cardiovascular events in patients suffering from an acute coronary syndrome. We sought to determine the impact of these 2 trials on trends in intensive versus moderate statin therapy in Ontario, Canada.

Methods and Results—

This is a retrospective time-series analysis of statin prescribing between June 1997 and September 2004 in Ontario, Canada, for all residents age 65 years and older. The publication of the PROVE IT–TIMI 22 and REVERSAL trials was associated with a sustained and statistically significant increase in the number of prescriptions dispensed for atorvastatin 80 mg (range, 272 to 635 additional prescriptions per month), whereas the number of prescriptions filled for pravastatin 40 mg did not change. Similarly, it resulted in a temporal increase in the relative market share of atorvastatin at a dose of 80 mg versus that of atorvastatin at a dose of 40 mg. However, the proportion of simvastatin prescriptions for 80 mg relative to 40 mg did not change over time, implying a drug-specific effect rather than a class effect in prescribing practice.

Conclusions—

The publication of the PROVE IT–TIMI 22 and REVERSAL trials resulted in a significant sustained increase in the use of intensive compared with moderate statin therapy. This shift was evident primarily in an increased use of high-dose atorvastatin and did not appear to be generalizable to other statins.
Endogenous Ouabain
Authors: Murrell Julie R. , Randall Jeffrey D. , Rosoff James , Zhao Ji-liang , Jensen Roderick V. , Gullans Steven R. , Haupert Garner T.
Published: August 2005

Background—

Mammalian tissues contain a presumed endogenous Na
+
, K
+
-ATPase inhibitor that binds reversibly to the Na
+
pump with high affinity and specificity. The inhibitor has been linked to the pathogenesis of experimental volume-expanded and human essential hypertension. This compound has been isolated from mammalian hypothalamus and appears to be an isomer of the plant-derived cardiac glycoside ouabain, if not ouabain itself. The objective of this study was to test the hypothesis that a biosynthetic pathway exists in mammalian tissues to produce a steroid derivative closely related to plant cardiac glycosides.

Methods and Results—

Using bioinformatics and genomic techniques, Milan hypertensive rat tissues were studied because this strain has a 10-fold increase in hypothalamic ouabain-like compound that is linked to the pathogenesis of the hypertension. A putative steroid biosynthetic pathway was constructed and candidate genes encoding enzymes in this pathway were identified from sequence databases. Differential expression of selected genes in the pathway was studied by microarray analysis and quantitative polymerase chain reaction, with functional validation by gene silencing using small interfering RNAs. Marked upregulation of genes coding for P450 side chain cleavage and Δ5-3β-hydroxysteroid dehydrogenase/Δ5-Δ4- isomerase enzymes in hypertensive hypothalamus but not adrenal was found, compared with normotensive Milan rats. Knockdown of the latter gene decreased production of ouabain-like factor from neural tissue.

Conclusions—

Our findings support the possibility that a unique steroid biosynthetic circuit exists in Milan rat brain, functioning independently from adrenal, which could account for the overproduction of the hypothalamic ouabain-like compound in this species.
Individualism-Collectivism and the Attitudes to School Bullying of Japanese and Australian Students
Authors: Nesdale Drew , Naito Mikako
Published: August 2005

This study examined whether collectivism versus individualism influences participants’ attitudes toward group-based bullying. Student members of a collectivistic culture (Japan; n = 158) versus an individualistic culture (Australia; n = 157) read about a school bullying episode. Collectivistic responses were predicted when the victim was a student from the same culture as the participant, and the classroom group had a norm of bullying versus helping. Individualistic responses were predicted when the participant learned that he or she was personally connected with the bully or the victim. Contrary to predictions, the participants’ attitudes reflected the interaction of nationality and gender, with the gender difference being greater between the Japanese versus Australian participants. In contrast, the participants’ behavior intentions mainly reflected their nationality—the Japanese participants revealed a greater likelihood of bullying, and a lower likelihood of helping a victim, than did the Australians. The implications for research on individualism-collectivism and bullying are discussed.
The Equivalence of the Behavior Problem Index Across U.S. Ethnic Groups
Authors: Spencer Michael S. , Fitch Dale , Grogan-Kaylor Andrew , Mcbeath Bowen
Published: August 2005

In this study, the authors examine the equivalence of the factor structure of a commonly used symptom checklist of behavioral and emotional problems—the Behavior Problem Index (BPI)—across African American, Hispanic, and White children in the United States. The sample is drawn from the 1998 data file of the Children of the National Longitudinal Survey of Youth, a U.S. data set. The results of the study suggest that the BPI is not equivalent across the three ethnic groups. These findings are consistent when equivalence is tested for a one-factor model, a two-factor model using the internalizing and externalizing dimensions of the BPI, and a six-factor model using the subscales of the BPI. Item-level analyses identify the statistically significant items that are associated with nonequivalence across ethnic groups. The implications of nonequivalent measures for cross-cultural research and practice with families and children are discussed.
Predicting Expatriate Job Performance for Selection Purposes
Authors: Mol Stefan T. , Born Marise Ph. , Willemsen Madde E. , Van Der Molen Henk T.
Published: August 2005

This article meta-analytically reviews empirical studies on the prediction of expatriate job performance. Using 30 primary studies (total N = 4,046), it was found that predictive validities of the Big Five were similar to Big Five validities reported for domestic employees. Extraversion, emotional stability, agreeableness, and conscientiousness were predictive of expatriate job performance; openness was not. Other predictors that were found to relate to expatriate job performance were cultural sensitivity and local language ability. Cultural flexibility, selection board ratings, tolerance for ambiguity, ego strength, peer nominations, task leadership, people leadership, social adaptability, and interpersonal interest emerged as predictors from exploratory investigations (K < 4). It is surprising that intelligence has seldom been investigated as a predictor of expatriate job performance.
Chinese and English Probabilistic Thinking and Risk Taking in Gambling
Authors: Lau Lai-Yin , Ranyard Rob
Published: August 2005

This study investigated cultural differences in probabilistic thinking and risk taking. A View of Uncertainty Questionnaire assessed the probabilistic thinking of Chinese and English gamblers and nongamblers and a hypothetical Horse Racing Task assessed risk-taking behavior ( N = 120). The Chinese exhibited significantly less probabilistic thinking and riskier gambling decisions. A path analysis found that degree of probabilistic thinking was one (but not the only) source of the observed cultural differences in risk taking.
Prospects for the UK Economy
Authors: Barrell Ray , Choy Amanda , Kirby Simon , Metz Robert , Pomerantz Olga
Published: August 2005

UK quarterly national accounts have undergone a rebasing to 2002 prices, together with a comprehensive revision to past data. The revised data paint a significantly different picture of the UK economy from that on which we based our last forecast. It now seems that the economy decelerated in the second half of 2004 and into the first quarter of 2005 (see chart 1), whereas the previous data indicated a more robust outcome for the second half of 2004, with greater momentum carrying over into 2005. The preliminary estimate of GDP suggests that the belowtrend growth continued into the second quarter of this year. The previous data indicated that the UK was operating at full capacity, while it would now seem that we are moving into a period where there is some spare capacity in the economy.
Extended Correlation
Authors: Levine Joel H.
Published: August 2005

What is the correlation between two variables? Traditional answers offer summary assessments such as Pearson’s r and regression coefficients. But new computing techniques make it possible to construct conceptually simple hypotheses that describe the full joint distribution of two variables, making it possible to “mine” the correlation for information that was previously unused. This article begins with evidence of systematic anomalies in the empirical joint distribution of height-weight data and follows with a hypothesis that explains these anomalies in terms of a theoretical joint distribution relative to a linear equation. The hypothesis has serious consequences because even in traditional examples, while it offers an improved fit to the data, its estimates of the linear center do not correspond to traditional least squares estimates of the linear relation for the same two variables.
The Impact of Community-Level Variables on Individual-Level Outcomes
Authors: Angeles Gustavo , Guilkey David K. , Mroz Thomas A.
Published: August 2005

The authors study alternative estimators of the impacts of higher level variables in multilevel models. This is important since many of the important variables in social science research are higher level factors having impacts on many lower level outcomes such as school achievement and contraceptive use. While the large sample properties of alternative estimators for these models are well known, there is little evidence about the relative performance of these estimators in the sample sizes typical in social science research. The authors attempt to fill this gap by presenting evidence about point estimation and standard error estimation for both two-and three-level models. A major conclusion of the article is that readily available commercial software can be used to obtain both reliable point estimates and coefficient standard errors in models with two or more levels as long as appropriate corrections are made for possible error correlations at the highest level.
PAIS: paracetamol (acetaminophen) in stroke; protocol for a randomized, double blind clinical trial. [ISCRTN 74418480]
Authors: van Breda Eric J , van der Worp H Bart , van Gemert H Maarten A , Algra Ale , Kappelle L Jaap , van Gijn Jan , Koudstaal Peter J , Dippel Diederik WJ ,   the PAIS investigators
Published: August 2005

Background

In patients with acute stroke, increased body temperature is associated with large lesion volumes, high case fatality, and poor functional outcome. A 1°C increase in body temperature may double the odds of poor outcome. Two randomized double-blind clinical trials in patients with acute ischemic stroke have shown that treatment with a daily dose of 6 g acetaminophen (paracetamol) results in a small but rapid and potentially worthwhile reduction of 0.3°C (95% CI: 0.1–0.5) in body temperature. We set out to test the hypothesis that early antipyretic therapy reduces the risk of death or dependency in patients with acute stroke, even if they are normothermic.

Methods/design

Paracetamol (Acetaminophen) In Stroke (PAIS) is a randomized, double-blind clinical trial, comparing high-dose acetaminophen with placebo in 2500 patients. Inclusion criteria are a clinical diagnosis of hemorrhagic or ischemic stroke and the possibility to start treatment within 12 hours from onset of symptoms. The study will have a power of 86% to detect an absolute difference of 6% in the risk of death or dependency at three months, and a power of 72% to detect an absolute difference of 5%, at a 5% significance level.

Discussion

This is a simple trial, with a drug that only has a small effect on body temperature in normothermic patients. However, when lowering body temperature with acetaminophen does have the expected effectiveness, 20 patients will have to be treated to prevent dependency or death in one.
Psoriasis vulgaris flare during efalizumab therapy does not preclude future use: a case series
Authors: Lowes Michelle A , Turton James A , Krueger James G , Barnetson Ross StC
Published: August 2005

Background

Severe psoriasis vulgaris can be extremely difficult to treat in some patients, even with the newer biological therapies available today.

Case presentations

We present two patients with severe chronic plaque psoriasis who received numerous systemic anti-psoriatic therapies with varied results. Both responded well to initial treatment with efalizumab (anti-CD11a), but then experienced a flare of their disease after missing a dose. However, after disease stablization, both patients responded well to re-introduction of efalizumab, one patient requiring concurrent treatment with infliximab (anti-TNF-α).

Conclusion

These cases are presented to characterize this "flare" reaction, and to inform health care providers that efalizumab can still be administered after disease flare, and again may be a successful therapy.
Effect of the degree of ischaemic injury and reoxygenation time on the type of myocardial cell death in man: role of caspases
Authors: Vohra Hunaid A , Galiñanes Manuel
Published: August 2005

Background

The importance of apoptosis in the injury sustained by the human myocardium during ischaemia and reoxygenation and the underlying mechanisms remain unclear. To quantify apoptosis and necrosis induced by simulated ischaemia/reoxygenation in the human atrial myocardium, free-hand sections of right atrial appendage (n = 8/group) were subjected to 90 minutes simulated ischaemia followed by 2, 8 and 24 hours reoxygenation.

Results

Apoptosis, as assessed by TUNEL, was greater than necrosis after 90 minutes simulated ischaemia and 2 hours reoxygenation (35.32 ± 3.22% vs 13.55 ± 1.3%; p < 0.05) but necrosis was greater than apoptosis by 24 hours reoxygenation (45.20 ± 2.75% vs 4.82 ± 0.79%; p < 0.05). Total caspase activation was similar after 90 minutes simulated ischaemia followed by 2 hours and 24 hours reoxygenation (515270 ± 99570 U vs 542940 ± 95216 U; p = NS). However, caspase-3 like activation was higher at 2 hours than at 24 hours reoxygenation (135900 ± 42200 U vs 54970 ± 19100 U; p < 0.05). Inhibition of caspase-3 by z.DEVD.fmk (70 nM) almost completely abolished apoptosis from 23.26 ± 2.854% to 0.73 ± 0.28 % (p < 0.05), without affecting necrosis.

Conclusion

Cell death by apoptosis and necrosis in the human myocardium subjected to simulated ischaemia/reoxygenation depends on the degree of the ischaemic insult and have a different time-course with apoptosis happening early during reoxygenation and necrosis becoming more important later. Importantly, the apoptosis induced by simulated ischaemia/reoxygenation is mainly mediated by activation of caspase-3 but it does not affect necrosis.
CXCR4 and CCR5 shRNA transgenic CD34+ cell derived macrophages are functionally normal and resist HIV-1 infection
Authors: Anderson Joseph , Akkina Ramesh
Published: August 2005

Background

Stable simultaneous knock down of the HIV-1 coreceptors CCR5 and CXCR4 is a promising strategy to protect cells from both R5 macrophage tropic and X4 T cell tropic as well as dual tropic viral infections. The potency of shRNAs in targeted gene silencing qualifies them as powerful tools for long term HIV gene therapy. Our previous work with a bispecific lentiviral vector containing CXCR4 and CCR5 shRNAs showed efficacy in down regulating both coreceptors and conferring viral resistance to both X4 and R5-tropic strains of HIV-1 in cultured cell lines. To extend these results to a stem cell gene therapy setting, here we show transduction of primary CD34+ hematopoietic progenitor cells to derive normal end stage cells that are resistant to HIV-1 infection.

Results

The bispecific XHR lentiviral vector harboring CXCR4 and CCR5 shRNA expression cassettes was efficient in transducing CD34+ cells. The transduced cells gave rise to morphologically normal transgenic macrophages when cultured in cytokine media. There was a marked down regulation of both coreceptors in the stably transduced macrophages which showed resistance to both R5 and X4 HIV-1 strains uponin vitrochallenge. Since off target effects by some shRNAs may have adverse effects on transgenic cells, the stably transduced macrophages were further analyzed to determine if they are phenotypically and functionally normal. FACS evaluation showed normal levels of the characteristic surface markers CD14, CD4, MHC class II, and B7.1. Phagocytic functions were also normal. The transgenic macrophages demonstrated normal abilities in up-regulating the costimulatory molecule B7.1 upon LPS stimulation. Furthermore, IL-1 and TNFα cytokine secretion in response to LPS stimulation was also normal. Thus, the transgenic macrophages appear to be phenotypically and functionally normal.

Conclusion

These studies have demonstrated for the first time that a bispecific lentiviral vector could be used to stably deliver shRNAs targeted to both CCR5 and CXCR4 coreceptors into CD34+ hematopoietic progenitor cells and derive transgenic macrophages. Transgenic macrophages with down regulated coreceptors were resistant to both R5 and X4 tropic HIV-1 infections. The differentiated cells were also phenotypically and functionally normal indicating no adverse effects of shRNAs on lineage specific differentiation of stem cells. It is now possible to construct gene therapeutic lentiviral vectors incorporating multiple shRNAs targeted to cellular molecules that aid in HIV-1 infection. Use of these vectors in a stem cell setting shows great promise for sustained HIV/AIDS gene therapy.
Prolonged Membrane Potential Depolarization in Cingulate Pyramidal Cells after Digit Amputation in Adult Rats
Authors: Wu MF , Pang ZP , Zhuo M , Xu ZC
Published: August 2005

The anterior cingulate cortex (ACC) plays an important role in higher brain functions including learning, memory, and persistent pain. Long-term potentiation of excitatory synaptic transmission has been observed in the ACC after digit amputation, which might contribute to plastic changes associated with the phantom pain. Here we report a long-lasting membrane potential depolarization in ACC neurons of adult rats after digit amputation in vivo. Shortly after digit amputation of the hind paw, the membrane potential of intracellularly recorded ACC neurons quickly depolarized from ∼−70 mV to ∼−15 mV and then slowly repolarized. The duration of this amputation-induced depolarization was about 40 min. Intracellular staining revealed that these neurons were pyramidal neurons in the ACC. The depolarization is activity-dependent, since peripheral application of lidocaine significantly reduced it. Furthermore, the depolarization was significantly reduced by a NMDA receptor antagonist MK-801. Our results provide direct in vivo electrophysiological evidence that ACC pyramidal cells undergo rapid and prolonged depolarization after digit amputation, and the amputation-induced depolarization in ACC neurons might be associated with the synaptic mechanisms for phantom pain.
Antifactor Xa activity in critically ill patients receiving antithrombotic prophylaxis with standard dosages of certoparin: a prospective, clinical study
Authors: Jochberger Stefan , Mayr Viktoria , Luckner Günter , Fries Dietmar R , Mayr Andreas J , Friesenecker Barbara E , Lorenz Ingo , Hasibeder Walter R , Ulmer Hanno , Schobersberger Wolfgang , Dünser Martin W
Published: August 2005

Introduction

Deep venous thrombosis with subsequent pulmonary embolism or post-thrombotic syndrome is a feared complication in the intensive care unit. Therefore, routine prophylactic anticoagulation is widely recommended. Aside from unfractionated heparin, low molecular weight heparins, such as certoparin, have become increasingly used for prophylactic anticoagulation in critically ill patients. In this prospective study, we evaluated the potency of 3,000 IU certoparin administered once daily to reach antithrombotic antifactor Xa (aFXa) levels of 0.1 to 0.3 IU/ml in 62 critically ill patients.

Methods

AFXa levels were determined 4, 12 and 24 h after injection of certoparin. Prothrombin time, activated partial thromboplastin time, antithrombin, fibrinogen, hemoglobin, platelet count, serum urea and creatinine concentrations were documented before and 12 and 24 h after injection of certoparin.

Results

Four hours after certoparin injection (n = 32), 28% of patients were within the antithrombotic aFXa range. After 12 and 24 h, 6% achieved antithrombotic aFXa levels. Because of a severe pulmonary embolism in one study patient, an interim analysis was performed, and the dosage of certoparin was increased to 3,000 IU twice daily. This regime attained recommended antithrombotic aFXa levels in 47%, 27%, 40% and 30% of patients at 4, 12, 16 and 24 h, respectively, after twice daily certoparin injection (n = 30). Antithrombin and fibrinogen concentrations slightly increased during the observation period. Low antithrombin concentrations before certoparin were independently correlated with underdosing of certoparin. Patients with aFXa levels <0.1 IU/ml 4 h after certoparin injection required vasopressors more often and had lower serum concentrations of creatinine and urea than patients with antithrombotic aFXa levels.

Conclusion

Standard dosages of certoparin of 3,000 IU given once or twice daily are ineffective for attaining the recommended aFXa levels of 0.1 to 0.3 IU/ml in critically ill patients. Low antithrombin levels before certoparin administration were independently associated with low aFXa levels. Renal function and vasopressor therapy may further influence the effectiveness of certoparin in ensuring adequate antithrombotic prophylaxis.
Does cardiac surgery in newborn infants compromise blood cell reactivity to endotoxin?
Authors: Schumacher Kathrin , Korr Stefanie , Vazquez-Jimenez Jaime F , von Bernuth Götz , Duchateau Jean , Seghaye Marie-Christine
Published: August 2005

Introduction

Neonatal cardiac surgery is associated with a systemic inflammatory reaction that might compromise the reactivity of blood cells against an inflammatory stimulus. Our prospective study was aimed at testing this hypothesis.

Methods

We investigated 17 newborn infants with transposition of the great arteries undergoing arterial switch operation. Ex vivo production of the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α), of the regulator of the acute-phase response IL-6, and of the natural anti-inflammatory cytokine IL-10 were measured by enzyme-linked immunosorbent assay in the cell culture supernatant after whole blood stimulation by the endotoxin lipopolysaccharide before, 5 and 10 days after the operation. Results were analyzed with respect to postoperative morbidity.

Results

The ex vivo production of TNF-α and IL-6 was significantly decreased (P < 0.001 and P < 0.002, respectively), whereas ex vivo production of IL-10 tended to be lower 5 days after the operation in comparison with preoperative values (P < 0.1). Ex vivo production of all cytokines reached preoperative values 10 days after cardiac surgery. Preoperative ex vivo production of IL-6 was inversely correlated with the postoperative oxygenation index 4 hours and 24 hours after the operation (P < 0.02). In contrast, postoperative ex vivo production of cytokines did not correlate with postoperative morbidity.

Conclusion

Our results show that cardiac surgery in newborn infants is associated with a transient but significant decrease in the ex vivo production of the pro-inflammatory cytokines TNF-α and IL-6 together with a less pronounced decrease in IL-10 production. This might indicate a transient postoperative anti-inflammatory shift of the cytokine balance in this age group. Our results suggest that higher preoperative ex vivo production of IL-6 is associated with a higher risk for postoperative pulmonary dysfunction.
Differential gene expression in anatomical compartments of the human eye
Authors: Diehn Jennifer J , Diehn Maximilian , Marmor Michael F , Brown Patrick O
Published: August 2005

Background

The human eye is composed of multiple compartments, diverse in form, function, and embryologic origin, that work in concert to provide us with our sense of sight. We set out to systematically characterize the global gene expression patterns that specify the distinctive characteristics of the various eye compartments.

Results

We used DNA microarrays representing approximately 30,000 human genes to analyze gene expression in the cornea, lens, iris, ciliary body, retina, and optic nerve. The distinctive patterns of expression in each compartment could be interpreted in relation to the physiology and cellular composition of each tissue. Notably, the sets of genes selectively expressed in the retina and in the lens were particularly large and diverse. Genes with roles in immune defense, particularly complement components, were expressed at especially high levels in the anterior segment tissues. We also found consistent differences between the gene expression patterns of the macula and peripheral retina, paralleling the differences in cell layer densities between these regions. Based on the hypothesis that genes responsible for diseases that affect a particular eye compartment are likely to be selectively expressed in that compartment, we compared our gene expression signatures with genetic mapping studies to identify candidate genes for diseases affecting the cornea, lens, and retina.

Conclusion

Through genome-scale gene expression profiling, we were able to discover distinct gene expression 'signatures' for each eye compartment and identified candidate disease genes that can serve as a reference database for investigating the physiology and pathophysiology of the eye.
Favorable Prognosis for Patients 12 to 18 Months of Age With Stage 4 Nonamplified MYCN Neuroblastoma: A Children's Cancer Group Study
Authors: Schmidt Mary Lou , Lal Ashutosh , Seeger Robert C. , Maris John M. , Shimada Hiroyuki , O'Leary Maura , Gerbing Robert B. , Matthay Katherine K.
Published: August 2005

Purpose

The long-term survival of children between age 12 and 24 months with stage 4 neuroblastoma and nonamplified MYCN (MYCN-NA) has not been defined previously.

Patients and Methods

Survival for stage 4 MYCN-NA neuroblastoma patients enrolled onto Children's Cancer Group (CCG) protocols 321P2 (1986 to 1991) and 3891 (1991 to 1996) was analyzed. Treatment consisted of intensive alkylator-based induction chemotherapy with or without autologous bone marrow transplantation (ABMT) with or without 13 cis-retinoic acid. Survival was analyzed by age strata less than 12, 12 to 18, 18 to 24, and more than 24 months at diagnosis. Patients younger than 12 months were treated on the moderate-intensity CCG protocol 3881.

Results

Forty-three patients with stage 4 MYCN-NA disease enrolled onto CCG-321P2 (n = 17) or CCG-3891 (n = 26) were between 12 and 24 months of age at diagnosis. After a median follow-up of 94 months (range, 4 to 140 months), the 6-year event-free survival (EFS) for the 12- to 18-month age group was superior to that of the 18- to 24-month age group (74% ± 8% v 31% ± 12%; P = .008). The EFS for children older than 24 months with stage 4 MYCN-NA neuroblastoma was 23% ± 3%, and for children younger than 12 months was 92% ± 3%.

Conclusion

Children diagnosed with stage 4 MYCN-NA neuroblastoma in the second year of life form a transitional group between infants and older children in terms of prognosis. Patients between 12 and 18 months of age have significantly better long-term survival than that of older children treated with intensive chemotherapy with or without ABMT. These patients may not benefit from additional intensification of therapy beyond that provided in earlier clinical trials and may even maintain this high survival rate with less intensive therapy.
Euthanasia and Depression: A Prospective Cohort Study Among Terminally Ill Cancer Patients
Authors: van der Lee Marije L. , van der Bom Johanna G. , Swarte Nikkie B. , Heintz A. Peter M. , de Graeff Alexander , van den Bout Jan
Published: August 2005

Purpose

To study the association between depression and the incidence of explicit requests for euthanasia in terminally ill cancer patients.

Patients and Methods

A prospective cohort study was conducted on 138 consecutive cancer patients with an estimated life expectancy of 3 months or less, in the period between September 1999 and August 2003. At inclusion, participants completed the Hospital Anxiety and Depression Scale. To identify “depressed mood” we used a cutoff score of 20. Kaplan-Meier curves and Cox regression analyses were used to assess the association between depressed mood and the risk of a request for euthanasia.

Results

Of 138 patients, 32 patients had depressed mood at inclusion. Thirty patients (22%) made an explicit request for euthanasia. The risk to request euthanasia for patients with depressed mood was 4.1 times higher than that of patients without depressed mood at inclusion (95% CI, 2.0 to 8.5).

Conclusion

Depression in cancer patients with an estimated life expectancy of less than 3 months is associated with a higher likelihood to request for euthanasia. The question of whether depressed mood can adequately be treated in this terminally ill population, and if so, whether it would lower the incidence of requests for euthanasia needs further investigation.
Expression of Serum Amyloid A, in Normal, Dysplastic, and Neoplastic Human Colonic Mucosa: Implication for a Role in Colonic Tumorigenesis
Authors: Gutfeld Orit , Prus Diana , Ackerman Zvi , Dishon Shira , Linke Reinhold P. , Levin Mark , Urieli-Shoval Simcha
Published: August 2005

Serum amyloid A (SAA) is an acute phase reactant, whose level in the blood is elevated in response to trauma, infection, inflammation, and neoplasia. Elevated levels of SAA in the serum of cancer patients were suggested to be of liver origin rather than a tumor cell product. The role of SAA in human malignancies has not been elucidated. We investigated the expression of SAA at various stages of human colon carcinoma progression. Nonradioactive in situ hybridization applied on paraffin tissue sections from 26 colon cancer patients revealed barely detected SAA mRNA expression in normal looking colonic epithelium. Expression was increased gradually as epithelial cells progressed through dysplasia to neoplasia. Deeply invading colon carcinoma cells showed the highest levels of SAA. Expression was also found in colon carcinoma metastases. Cells of lymphoid follicles of the intestinal wall, inflammatory cells, ganglion cells, and endothelial cells, also expressed SAA mRNA. Immunohistochemical staining revealed SAA protein expression that colocalized with SAA mRNA expression. RT-PCR analysis confirmed the expression of the SAA1 and SAA4 genes in colon carcinomas, expression that was barely detectable in normal colon tissues. These findings indicate local and differential expression of SAA in human colon cancer tissues and suggest its role in colonic tumorigenesis.
Conversions of Formaldehyde-modified 2′-Deoxyadenosine 5′-Monophosphate in Conditions Modeling Formalin-fixed Tissue Dehydration
Authors: Rait Vladimir K. , Zhang Qingrong , Fabris Daniele , Mason Jeffrey T. , O'Leary Timothy J.
Published: August 2005

Formalin-fixed, paraffin-embedded specimens typically provide molecular biologists with low yields of extractable nucleic acids that exhibit extensive strand cleavage and covalent modification of nucleic acid bases. This study supports the idea that these deleterious effects are promoted by the first step in formalin-fixed tissue processing—i.e., tissue dehydration with a graded series of alcohols. We analyzed the conversions of formaldehyde-modified 2′-deoxyadenosine 5′-monophosphate (dAMP) by reverse-phase ion-pair, high-performance liquid chromatography and found that dehydration does not stabilize N-methylol groups in the modified nucleotide. Furthermore, spontaneous demodification in a dry state or in anhydrous ethanol can be as fast as it is in aqueous solutions if the preparation is contaminated with salts of orthophosphoric acid. In ethanol, orthophosphates also catalyze formation of abundant N6-ethoxymethyl-dAMP, as well as cross-linking and depurination of nucleotides present in the mixture. Identification of the products was performed using ultraviolet absorbance spectroscopy and electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry. Alternatives to the traditional processing of formalin-fixed tissues are discussed.
Germinal Center Dendritic Cells Express More ICAM-1 Than Extrafollicular Dendritic Cells and ICAM-1/LFA-1 Interactions are Involved in the Capacity of Dendritic Cells to Induce PBMCs Proliferation
Authors: Goval Jean-Jacques , Greimers Roland , Boniver Jacques , de Leval Laurence
Published: August 2005

Germinal center dendritic cells (GCDCs) have been identified as CD11c+ CD4+ CD3− cells located in GCs with the ability of inducing marked proliferation of allogenic T cells. Using immunofluorescence techniques, we have observed that this CD11c+ CD4+ CD3− immunophenotype identified GCDCs but also a subset of extrafollicular DCs. By flow cytometry, we were able to discriminate the GCDCs (CD11chigh CD4high lin−) from the other tonsil DCs. By immunofluorescence and flow cytometry, we found that dendritic cells of germinal centers express more intracellular adhesion molecule-1 (ICAM-1) (CD54) than extrafollicular dendritic cells. Proliferation of peripheral blood mononuclear cells (PBMCs) induced by coculture with purified CD11c+ CD4+ CD3− DCs was reduced by addition of blocking anti-CD54 antibodies. In summary, distinct levels of ICAM-1 expression allow the distinction between GCDCs and extrafollicular DCs, and cellular interactions mediated by CD54 are likely to play a role in the capacity of GCDC to stimulate allogenic PBMC proliferation.
In Situ Localization of Transketolase Activity in Epithelial Cells of Different Rat Tissues and Subcellularly in Liver Parenchymal Cells
Authors: Boren Joan , Ramos-Montoya Antonio , Bosch Klazien S. , Vreeling Heleen , Jonker Ard , Centelles Josep J. , Cascante Marta , Frederiks Wilma M.
Published: August 2005

Metabolic mapping of enzyme activities (enzyme histochemistry) is an important tool to understand (patho)physiological functions of enzymes. A new enzyme histochemical method has been developed to detect transketolase activity in situ in various rat tissues and its ultrastructural localization in individual cells. In situ detection of transketolase is important because this multifunctional enzyme has been related with diseases such as cancer, diabetes, Alzheimer's disease, and Wernicke-Korsakoff's syndrome. The proposed method is based on the tetrazolium salt method applied to unfixed cryostat sections in the presence of polyvinyl alcohol. The method appeared to be specific for transketolase activity when the proper control reaction is performed and showed a linear increase of the amount of final reaction product with incubation time. Transketolase activity was studied in liver, small intestine, trachea, tongue, kidney, adrenal gland, and eye. Activity was found in liver parenchyma, epithelium of small intestine, trachea, tongue, proximal tubules of kidney and cornea, and ganglion cells in medulla of adrenal gland. To demonstrate transketolase activity ultrastructurally in liver parenchymal cells, the cupper iron method was used. It was shown that transketolase activity was present in peroxisomes and at membranes of granular endoplasmic reticulum. This ultrastructural localization is similar to that of glucose-6-phosphate dehydrogenase activity, suggesting activity of the pentose phosphate pathway at these sites. It is concluded that the method developed for in situ localization of transketolase activity for light and electron microscopy is specific and allows further investigation of the role of transketolase in (proliferation of) cancer cells and other pathophysiological processes.
The ATP-binding Cassette Transporter ABCG2 (BCRP), a Marker for Side Population Stem Cells, Is Expressed in Human Heart
Authors: Meissner Konrad , Heydrich Björn , Jedlitschky Gabriele , Schwabedissen Henriette Meyer zu , Mosyagin Igor , Dazert Peter , Eckel Lothar , Vogelgesang Silke , Warzok Rolf W. , Böhm Michael , Lehmann Christian , Wendt Michael , Cascorbi Ingolf , Kroemer Heyo K.
Published: August 2005

Efforts to improve severely impaired myocardial function include transplantation of autologous hematopoietic side population (SP) stem cells. The transmembrane ABC-type (ATP binding cassette) half-transporter ABCG2 (BCRP) serves as a marker protein for SP cell selection. We have recently shown that other ABC transport proteins such as ABCB1 and ABCC5 are differentially expressed in normal and diseased human heart. Here we investigated localization and individual ABCG2 expression in 15 ventricular (including 10 cardiomyopathic) and 51 auricular heart tissue samples using immunohistochemistry, confocal laser scanning fluorescence microscopy, and real-time RT-PCR. Individual genotypes were assigned using PCR–restriction fragment length polymorphism (RFLP) analysis and subsequently correlated to ABCG2 mRNA levels. ABCG2 was localized in endothelial cells of capillaries and arterioles of all samples. Ventricular samples from cardiomyopathic hearts exhibited significantly increased levels of ABCG2 mRNA (ABCG2/18S rRNA: 1.08 ± 0.30 × 10−7; p = 0.028 (dilative cardiomyopathy) and 1.16 ± 0.46 × 10−7; p = 0.009 (ischemic cardiomyopathy) compared with 0.44 ± 0.26 × 10−7 in nonfailing hearts). The individual haplotypes were not associated with altered mRNA expression. ABCG2 is variably expressed in endothelial cells of human heart, where it may function as a protective barrier against cardiotoxic drugs such as anthracyclines or mitoxantrone. ABCG2 expression is induced in dilative and ischemic cardiomyopathies.
Novel applications of the intraoperative parathyroid hormone assay Neue Anwendungen des intraoperativen Parathormon-Assays
Authors: Richards Melanie L. , Bingener Juliane
Published: August 2005

The development of an accurate and rapid method to measure parathyroid hormone (PTH) utilizing a two-site immunochemiluminometric assay (ICMA) has expanded its use to point-of-care testing. Measuring parathyroid hormone can be utilized to determine the completeness of parathyroid resection, to regionalize hypersecreting parathyroid glands, to identify parathyroid glands on fine-needle aspiration, and as a predictor of postoperative hypocalcemia following thyroidectomy and parathyroidectomy. This review addresses the current utilization of these techniques in patients with thyroid and parathyroid disease.