bluuebunny/crossref_metadata_2025 huggingface dataset:
Contrast-Enhanced Magnetic Resonance Angiography of Carotid Arteries
Authors: Johnston Dean C.C. , Eastwood James D. , Nguyen Thanh , Goldstein Larry B.
Published: December 2002

Background and Purpose—

Contrast-enhanced magnetic resonance angiography (CEMRA) is among the newer noninvasive tests used for the evaluation of patients with carotid artery disease. Evidence supporting its utility in routine clinical practice is lacking.

Methods—

The results of CEMRA were compared with those of catheter angiography in 50 consecutive patients being evaluated for carotid endarterectomy (CEA) at a community hospital. Using indications for CEA based on published guidelines, we determined the rate of misclassification for surgery, sensitivity, specificity, and positive and negative predictive values. In addition, the interrater agreement (κ score) of CEMRA was compared with that of catheter angiography in the studied population and with interpretations provided by 2 blinded radiologists.

Results—

Compared with catheter angiography, 24% (95% CI, 12% to 36%) of patients would have been misclassified for CEA on the basis of CEMRA results alone. CEMRA was associated with sensitivity of 92%, specificity of 62%, positive predictive value of 78%, and negative predictive value of 89%. When both CEMRA and duplex Doppler ultrasound were performed and the results were concordant, the misclassification rate decreased to 17% (95% CI, 2% to 32%). κ scores were similar for CEMRA and catheter angiography (0.72 and 0.75, respectively).

Conclusions—

CEMRA was found to be highly sensitive for detection of surgically amenable carotid stenosis. κ scores for the interpretation of CEMRA and catheter angiography were similar. However, clinicians should be cautious when using CEMRA alone for surgical decision making in CEA candidates because a significant number of patients may be misclassified. The rate of misclassification is reduced when the results of CEMRA and duplex Doppler ultrasound are concordant.
Carotid Endarterectomy and Race
Authors: Oddone Eugene Z. , Horner Ronnie D. , Johnston Dean C.C. , Stechuchak Karen , McIntyre Lauren , Ward Aileen , Alley Linda G. , Whittle Jeff , Kroupa Laura , Taylor John
Published: December 2002

Background and Purpose—

Carotid endarterectomy (CE) has been proved to reduce the risk of stroke for certain patients, but black patients are less likely than whites to receive CE. The purpose of this work was to determine the importance of clinical indications and patient preferences in predicting the use of carotid angiography and CE in a racially stratified sample of patients.

Methods—

Between 1997 and 1999, 708 patients with at least 1 carotid artery containing a ≥50% stenosis were enrolled (617 whites, 91 blacks) from 5 Veteran Affairs Medical Centers. Patient interviews were conducted at the time of the index carotid ultrasound, and each patient was followed up for 6 months to determine clinical events and receipt of carotid angiography or CE.

Results—

Black and white patients were similar in terms of age, sex, education level, and social support. More black than white patients received ultrasound for a completed stroke (36% versus 13%), and fewer black patients were classified as asymptomatic (56% versus 70%) or as having had a TIA (8% versus 17%;
P
<0.001). Health-related quality of life scores, trust in physician, and medical comorbidity scores were similar for black and white patients. Black patients expressed higher aversion to CE than white patients (31% versus 15% in the highest aversion quartile for blacks and whites, respectively;
P
=0.01). During follow-up, 20% of white patients and 14% of black patients received CE (
P
=0.19). In adjusted analyses, only patient clinical status as it relates to the indication for CE and site were associated with receipt of CE.

Conclusions—

Contrary to prior research, patient’s race was not associated with receipt of invasive carotid imaging or CE for older male veterans. These findings persist after controlling for patient preferences, comorbid illness, and quality of life. For patients enrolled in an equal-access healthcare system, clinical status was the primary determinant of the receipt of CE.
Endothelin-1–Induced Vasospasms of Spiral Modiolar Artery Are Mediated by Rho-Kinase–Induced Ca
2+
Sensitization of Contractile Apparatus and Reversed by Calcitonin Gene–Related Peptide
Authors: Scherer Elias Q. , Herzog Michael , Wangemann Philine
Published: December 2002

Background and Purpose—

Vasospasms of the spiral modiolar artery may cause an ischemic stroke of the inner ear that manifests itself by a sudden hearing loss. Previously we have shown that endothelin-1 (ET-1) induces vasospasms of the spiral modiolar artery. Here we tested the hypotheses that ET-1–induced vasospasms are (1) reversible by ET
A
receptor antagonists; (2) mediated by a Ca
2+
sensitization of the contractile apparatus via a Rho-kinase–induced inhibition of myosin light chain phosphatase; and (3) reversible by the vasodilator calcitonin gene–related peptide (CGRP).

Methods—

The Ca
2+
sensitivity of the contractile apparatus was evaluated by correlation between the smooth muscle cell Ca
2+
concentration and the vascular diameter, which were measured by microfluorometry with the fluorescent dye fluo-4 and videomicroscopy, respectively.

Results—

ET-1–induced vasospasms were prevented but not reversed by the ET
A
receptor antagonists BQ-123 and BMS-182874. The Ca
2+
sensitivity of the contractile apparatus was increased by ET-1 and by inhibition of myosin light chain phosphatase with calyculin A and was decreased by CGRP. ET-1–induced vasospasms and Ca
2+
sensitization were prevented and reversed by the Rho-kinase antagonist Y-27632 and by CGRP.

Conclusions—

ET-1 induces vasospasms of the spiral modiolar artery via ET
A
receptor–mediated activation of Rho-kinase, inhibition of myosin light chain phosphatase, and an increase in Ca
2+
sensitivity, which is reversed by CGRP. The observation that vasospasms were reversed by Y-27632 but not by BQ-123 or BMS-182874 suggests that Rho-kinase, rather than the ET
A
receptor, is the most promising pharmacological target for the treatment of ET-1–induced vasospasms, ischemic strokes, and sudden hearing loss.
A Population-Based Study of Brain Arteriovenous Malformation
Authors: ApSimon H.T. , Reef H. , Phadke R.V. , Popovic E.A.
Published: December 2002

Background and Purpose—

By undertaking long-term follow-up of a functionally isolated population study group, we sought to achieve a true picture of intrinsic brain arteriovenous malformation (BAVM). We sought to assess the validity of earlier population-based series and to determine the effects of newer treatment methods on the overall morbidity and mortality of BAVM.

Methods—

We excluded other intracranial vascular pathologies by defining criteria. By retrospective and prospective study, 240 patients with BAVM were followed for a mean of 10.11 years from first diagnosis.

Results—

Death rates were as follows: all causes, 12.9%; all BAVM related, 8.75%; BAVM related during conservative management, 24.6%; and BAVM related during active management, 3.9% (
P
=0.031). Mean diagnosis-to-death interval was 10.6 years. Oxford neurological disability scale grades of 209 survivors (July 2001) were as follows: grades 0 to 2, 74.1%; grade 3, 17.2%; and grades 4 to 5, 9.5%. Death rates were higher for patients who had bled or suffered nonhemorrhagic neurological deficit at original presentation. Incidence of first-ever hemorrhage in untreated patients was as follows: 0 to 9 years, 4.6% (
P
=0.0035); 30 to 39 years, 21% (
P
=0.02); and 60 to 69 years, 40.0% (
P
=0.045). The first bleed was fatal in 4.6%.

Conclusions—

We find no evidence of a substantial undiagnosed reservoir of nonsymptomatic BAVM. All BAVM are potentially hazardous. The great majority of BAVM patients become symptomatic during the patient’s lifetime, and the majority will bleed. The risk of first hemorrhage is lifelong and rises with age. Compared with earlier population-based series, our low overall patient mortality is predominantly due to higher proportions of active treatment in the 1980s and 1990s.
Long-Term Outcome After Angioplasty for Symptomatic Extracranial Carotid Stenosis in Poor Surgical Candidates
Authors: Fox Douglas J. , Moran Christopher J. , Cross DeWitte T. , Grubb Robert L. , Rich Keith M. , Chicoine Michael R. , Dacey Ralph G. , Derdeyn Colin P.
Published: December 2002

Background and Purpose—

The optimal treatment of patients with symptomatic carotid stenosis who are poor surgical candidates is uncertain. The purposes of this study were to report the long-term outcome after angioplasty in a series of these patients and to compare these data with historical control data from the North American Symptomatic Carotid Endarterectomy Trial (NASCET).

Methods—

We identified 42 consecutive patients with >70% carotid stenosis and ipsilateral ischemic symptoms within 120 days of treatment with angioplasty. All were considered poor surgical candidates by experienced surgeons. Baseline epidemiological stroke risk factors were obtained from review of medical records. Follow-up was from clinic records and by telephone.

Results—

Baseline epidemiological stroke risk factors were similar to those of medically treated NASCET patients. Angioplasty patients tended to have higher degrees of stenosis (45% with >90% stenosis versus 24% in NASCET) and more frequent contralateral stenosis or occlusion (30% versus 9%) than NASCET patients. Three patients suffered procedural strokes; 2 of the 3 made nearly complete recoveries. One additional patient suffered a central retinal occlusion 48 hours after angioplasty. No ipsilateral strokes occurred during the mean follow-up period of 1.7 years. Three patients were lost to follow-up. The cumulative risk of stroke was 9.5% (4 of 42) compared with 26% at 2 years for medically treated patients in NASCET.

Conclusions—

These pilot data suggest a beneficial effect of angioplasty for patients with high-grade symptomatic carotid stenosis who are not good surgical candidates.
AMPA Antagonist ZK200775 in Patients With Acute Ischemic Stroke
Authors: Elting Jan-Willem , Sulter Geert A. , Kaste Markku , Lees Kennedy R. , Diener Hans C. , Hommel Marc , Versavel Mark , Teelken Albert W. , De Keyser Jacques
Published: December 2002

Background and Purpose—

S-100B and neuron-specific enolase (NSE) serum concentrations can be used as peripheral markers of glial cell and neuronal damage, respectively. We investigated these markers in a clinical trial with the α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) antagonist ZK200775 in acute ischemic stroke patients.

Methods—

In a multicenter, double-blind, randomized, placebo-controlled phase 2 trial, 61 ischemic stroke patients were treated with either placebo or active drug in a dose-finding design. Twenty-five patients received placebo, 12 patients received a total dose of 262.5 mg in 48 hours (dose group 1), and 13 patients received a total dose of 525 mg in 48 hours (dose group 2). Eleven patients received a total dose of 105 mg over a period of 6 hours (dose group 3; reduction of total dose and infusion time because of adverse events in group 2). Serum concentrations of S-100B and NSE were analyzed with the use of a monoclonal sandwich immunoluminometric assay. Neurological outcome was assessed with the National Institutes of Health Stroke Scale (NIHSS).

Results—

In group 2 there was a significant transient worsening in the mean NIHSS score 48 hours after the start of treatment. The mean increase was 11 points. This was due to reduction of consciousness (stupor and coma) in 8 of 13 patients. Neurological deterioration in group 2 was associated with a higher increase of S-100B concentrations, but not of NSE concentrations, than in the placebo group. The trial was stopped prematurely for safety reasons.

Conclusions—

The AMPA antagonist ZK200775 transiently worsened the neurological condition in patients with acute ischemic stroke. Our results suggest that in addition to neuronal dysfunction, glial cell toxicity may have occurred. It may be useful to introduce monitoring of serum markers of brain damage in phase 2 trials with glutamate receptor antagonists.
Genetic Basis of Variation in Carotid Artery Plaque in the San Antonio Family Heart Study
Authors: Hunt Kelly J. , Duggirala Ravindranath , Göring Harald H.H. , Williams Jeff T. , Almasy Laura , Blangero John , O’Leary Daniel H. , Stern Michael P.
Published: December 2002

Background and Purpose—

In contrast to the commonly used quantitative marker of subclinical atherosclerosis, namely intima-media thickness, we investigated the extent to which the presence or absence of carotid artery plaque (CAP) was under genetic control.

Methods—

The study population consisted of 750 individuals distributed across 29 randomly ascertained extended Mexican American pedigrees who participated in the second examination cycle of the San Antonio Family Heart Study. Extracranial focal CAP was identified by B-mode ultrasound bilaterally in the internal carotid artery or the carotid bulb. Using a variance decomposition approach implemented in the SOLAR computer program, we performed genetic analysis on the discrete trait CAP (ie, liability to disease) using a threshold model. Covariates considered in the analysis included age, sex, diabetes, current smoking status, lipid levels, and markers of hypertension and obesity.

Results—

Fifty-one of 461 women and fifty-seven of 289 men with a mean age of 42.1 years had evidence of a plaque in the right and/or left carotid artery. The age- and sex-adjusted heritability (
h
2
±SE) for CAP was significant (
h
2
=0.28±0.15,
P
=0.01). Furthermore, after adjustment for additional covariates that contributed significantly to the model (
P
<0.05; diabetes, hypertension, body mass index, waist circumference, and smoking status), heritability remained significant (
h
2
=0.23±0.15,
P
=0.03).

Conclusions—

Our data indicate that after established cardiovascular risk factors are controlled for, the variation of the discrete trait CAP is under appreciable additive genetic influences.
Shortening the NIH Stroke Scale for Use in the Prehospital Setting
Authors: Tirschwell David L. , Longstreth W.T. , Becker Kyra J. , Gammans Richard E. , Sabounjian LuAnn A. , Hamilton Scott , Morgenstern Lewis B.
Published: December 2002

Background and Purpose—

Prehospital stroke scales should identify stroke patients and measure stroke severity. The goal of this study was to identify a subset of the 15 items in the National Institutes of Health Stroke Scale (NIHSS-15) that measures stroke severity and predicts outcomes.

Methods—

Using 2 distinct data sets from acute stroke clinical trials, we derived and validated shortened versions of the NIHSS (sNIHSS). Stepwise logistic regression and bootstrap techniques were used in selection of NIHSS-15 items. Areas under the receiver operator characteristic curve (C statistics) were used to compare predictive performance of logistic models incorporating differing versions of the NIHSS.

Results—

The derivation analyses suggested the 8 NIHSS-15 items that were most predictive of “good outcome” 3 months after stroke, in order of decreasing importance: right leg item, left leg, gaze, visual fields, language, level of consciousness, facial palsy, and dysarthria. The sNIHSS-8 comprises all 8 and the sNIHSS-5, the first 5. In the validation models, C statistics were NIHSS-15=0.80, sNIHSS-8=0.77, and sNIHSS-5=0.76. Statistical comparisons suggested that the NIHSS-15 had better predictive performance than the sNIHSS-8 or the sNIHSS-5; the absolute difference in C statistics was small. There was no significant difference between the sNIHSS-8 and the sNIHSS-5.

Conclusions—

Much of the predictive performance of the full NIHSS-15 was retained with a shortened scale, the sNIHSS-5. Shortening the NIHSS-15 will facilitate its use during prehospital evaluations. The sNIHSS severity information may be useful to triage acute stroke patients in communities and to provide a baseline stroke severity for prehospital acute stroke trials.
C-Reactive Protein Predicts Progression of Atherosclerosis Measured at Various Sites in the Arterial Tree
Authors: van der Meer Irene M. , de Maat Moniek P.M. , Hak A. Elisabeth , Kiliaan Amanda J. , del Sol Antonio Iglesias , van der Kuip Deirdre A.M. , Nijhuis Rogier L.G. , Hofman Albert , Witteman Jacqueline C.M.
Published: December 2002

Background and Purpose—

C-reactive protein (CRP) predicts myocardial infarction and stroke. Its role as a predictor of the progression of subclinical atherosclerosis is not yet known. We investigated whether CRP predicts progression of atherosclerosis measured at various sites in the arterial tree.

Methods—

CRP levels were measured in a random sample of 773 subjects ≥55 years of age who were participating in the Rotterdam Study. Subclinical atherosclerosis was assessed at various sites at 2 points in time, with a mean duration between measurements of 6.5 years.

Results—

After adjustment for age, sex, and smoking habits, odds ratios (ORs) associated with CRP levels in the highest compared with the lowest quartile were increased for progression of carotid (OR, 1.9; 95% CI, 1.1 to 3.3), aortic (OR, 1.7; 95% CI, 1.0 to 3.0), iliac (OR, 2.0; 95% CI, 1.2 to 3.3), and lower extremity (OR, 1.9; 95% CI, 1.0 to 3.7) atherosclerosis. The OR for generalized progression of atherosclerosis as indicated by a composite progression score was 4.5 (95% CI, 2.3 to 8.5). Except for aortic atherosclerosis, these estimates hardly changed after additional adjustment for multiple cardiovascular risk factors. In addition, ORs for progression of atherosclerosis associated with high CRP levels were as high as those associated with the traditional cardiovascular risk factors high cholesterol, hypertension, and smoking. Geometric mean levels of CRP increased with the total number of sites showing progression of atherosclerosis (
P
=0.002 for trend).

Conclusions—

CRP predicts progression of atherosclerosis measured at various sites in the arterial tree.
Gross Variability in the Detection of Prolactin in Sera Containing Big Big Prolactin (Macroprolactin) by Commercial Immunoassays
Authors: Smith Thomas P. , Suliman Abdulwahab M. , Fahie-Wilson Michael N. , McKenna T. Joseph
Published: December 2002

A high molecular mass form of prolactin (PRL), macroprolactin, accumulates in the sera of some subjects. Although macroprolactin exhibits limited bioactivity in vivo, it retains immunoreactivity. We examined the frequency of macroprolactinemia in clinical practice and the ability of immunoassay systems to distinguish between macroprolactin and monomeric PRL. Of 300 hyperprolactinemic sera identified, 71 normalized following treatment of sera with polyethylene glycol, indicating that 24% of hyperprolactinemia could be accounted for by macroprolactin. Ten of these macroprolactinemic sera were circulated to 18 clinical laboratories. Two sets of PRL measurements of the 10 untreated sera were obtained from each of the nine most commonly used immunoassay systems. Across the nine assay systems, differences in the PRL estimates ranged from 2.3- to 7.8-fold. Elecsys users reported the highest PRL levels. Somewhat lower values were reported for DELFIA systems followed by Immuno-1, AxSYM, and Architect assays. The Immulite 2000 assay generated PRL levels equivalent to approximately 50% of those reported by the high-reading methods. The lowest PRL levels were reported by Access, ACS:180, and Centaur systems. To avoid confusion caused by the frequent presence of macroprolactin accounting for hyperprolactinemia, secondary screening for the presence of macroprolactin is recommended.
Estradiol and Interleukin-1β Exert a Synergistic Stimulatory Effect on the Expression of the Chemokine Regulated upon Activation, Normal T Cell Expressed, and Secreted in Endometriotic Cells
Authors: Akoum Ali , Lemay André , Maheux Rodolphe
Published: December 2002

Endometriosis, commonly associated with intraperitoneal inflammation, is estrogen dependent. Possible links between the immunoinflammatory and endocrine changes observed in endometriotic women have been poorly understood. In this study, we report that estradiol (E2) and IL-1β exert a synergistic stimulatory action on RANTES (regulated upon activation, normal T cell expressed, and secreted) expression by endometriotic cells. Treatment of endometriotic cells with IL-1β had a dose-dependent effect on RANTES protein secretion and mRNA steady state levels, whereas cell treatment with E2 or progesterone had no detectable effect. Interestingly, treatment of endometriotic cells with E2 before stimulation with IL-1β resulted in a further increase in RANTES protein secretion and mRNA steady state levels, compared with IL-1β alone, whereas treatment with progesterone did not significantly affect cell responsiveness to IL-1β. Assessment of RANTES mRNA half-life revealed that cell pretreatment with E2 enhanced RANTES mRNA stability and increased gene transcription as shown by run-on analysis. Immunohistochemical analysis of RANTES in endometriotic tissue showed immunostaining to be predominant in the stroma with no noticeable differences in tissues from the proliferative and secretory phase of the menstrual cycle. This appears to be consistent with the cell culture data and indicates that RANTES expression in endometriotic tissue is not subject to cyclic variation. These findings reveal a new regulatory mechanism by which IL-1β produced by activated macrophages can in synergy with ovarian and locally produced E2 lead to enhanced macrophage and T-lymphocyte recruitment, thereby exacerbating the local immunoinflammatory process. Furthermore, the findings provide a further evidence for a close relationship between the endocrine and immunological changes observed in endometriosis.
Neurosurgical Treatment of Nelson’s Syndrome
Authors: Kelly P. A. , Samandouras G. , Grossman A. B. , Afshar F. , Besser G. M. , Jenkins P. J.
Published: December 2002

Total bilateral adrenalectomy remains the definitive procedure for cure in Cushing’s disease. It is complicated by the development of Nelson’s syndrome, the treatment of which remains troublesome. We report the long-term follow-up, median 17 yr (range, 8–22 yr), of 13 patients (3 males and 10 females) treated with pituitary surgery for Nelson’s syndrome at a median age of 35 yr (range, 21–67 yr). The presence of a pituitary mass lesion necessitated neurosurgery in all. Preoperatively, the median plasma ACTH level was 664 pmol/liter (range, 92–3665 pmol/liter); this fell to 29 pmol/liter (range, <2 to 1124 pmol/liter) postoperatively (P < 0.0005). Cutaneous hyperpigmentation was reduced in all and resolved in 11 patients. The pituitary tumor bulk was clearly reduced in 12 patients. There was no perioperative mortality. No patient developed a visual field defect attributable to surgery. New anterior pituitary hormone deficiency occurred in seven patients, and permanent diabetes insipidus occurred in five patients. At latest follow-up, the median plasma ACTH is 73 pmol/liter (range, <2 to 7759 pmol/liter); six patients have levels of less than 44 pmol/liter and also have a resolution of their pigmentation and no evidence of a recurrence of a pituitary mass lesion. We conclude that pituitary surgery is an efficacious treatment for mass lesions associated with Nelson’s syndrome, has long-term benefit with minimal side effects, and must be considered in the management of this distressing complication.
Small Guanosine Triphospatase RhoA and Rho-Associated Kinase as Regulators of Trophoblast Migration
Authors: Shiokawa Shigetatsu , Iwashita Mitsutoshi , Akimoto Yoshihiro , Nagamatsu Shinya , Sakai Ken , Hanashi Hideki , Kabir-Salmani Maryam , Nakamura Yukio , Uehata Masayoshi , Yoshimura Yasunori
Published: December 2002

The small guanosine triphosphatase Rho controls cell adhesion and motility through reorganization of the actin cyto-skeleton and regulation of actomyosin contractility. Among the putative target molecules of Rho, a Rho-associated coiled coil-forming protein kinase (ROCK) is thought to participate in Rho-mediated cell adhesion and motility. In the present study, we explored the expression and function of RhoA and ROCK in human trophoblast cells. The colocalization of RhoA, cytokeratin 8/18, and cytokeratin 7 in some cells located in the decidual stromal region indicated that extravillous trophoblast cells expressed RhoA. In double staining for RhoA and ROCK in human chorionic villi, RhoA staining was strongly positive in the cytoplasm of cytotrophoblasts, whereas ROCK stained in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts. Both RhoA and ROCK were stained in cytoplasma of cultured human cytotrophoblast. Cultured human trophoblast cells contained actin stress fibers that were lost after treatment with C3, an exoenzyme produced by Clostridium botulinum. Y-27632, a selective ROCK inhibitor, suppressed RhoA-induced formation of actin stress fibers and formation of focal contact in trophoblast cells. The trophoblast reacquired actin stress fibers and focal contact after withdrawal of Y-27632. Cultured human cytotrophoblast cells from 7–9 wk of gestation migrated into a fibronectin-coated membrane. Both C3 exoenzyme and Y-27632 inhibited cytotrophoblast migration in a dose-dependent manner. In conclusion, cyto-trophoblasts express RhoA and ROCK in their cytoplasm, and RhoA-ROCK is involved in their assembly of actin stress fibers. Suppression of RhoA-ROCK reduces trophoblast migration. These findings suggest that RhoA-ROCK signaling is a key regulator of trophoblast cell migration.
Characterization of Morphological and Cytoskeletal Changes in Trophoblast Cells Induced by Insulin-Like Growth Factor-I
Authors: Kabir-Salmani Maryam , Shiokawa Shigetatsu , Akimoto Yoshihiro , Hasan-Nejad Habib , Sakai Keiji , Nagamatsu Shinya , Sakai Ken , Nakamura Yukio , Hosseini Ahmad , Iwashita Mitsutoshi
Published: December 2002

IGF-I and IGF-II were appeared to play major roles in the adhesive and migratory events that are considered to be crucial in the implantation process. The purpose of this study was to determine the effects of IGF-I on trophoblast adhesion to extracellular matrix. Trophoblast cells obtained from early gestation at artificial abortion were incubated with the indicated doses of IGF-I at the indicated times. Trophoblast cells were treated with IGF-I in the presence or absence of RGD peptide and an antibody against α-subunit of IGF-I receptor (αIR3). Morphometric and morphological changes were studied using light and electron microscopy. Furthermore, vinculin, actin stress fibers, phosphorylated focal adhesion kinase (FAK), phosphotyrosine, and paxillin were immunolocalized in trophoblast cells after IGF-I treatment in the presence or absence of αIR3. Immunoprecipitation and anti-phosphotyrosine immunoblotting were carried out to detect the phosphorylated FAK and phosphorylated paxillin contents of the IGF-I-treated and untreated trophoblast cells. The results showed that IGF-I promoted trophoblast adhesion to fibronectin substrate in a time- and dose-dependent manner, and addition of RGD peptide and αIR3 monoclonal antibody abolished the effects of IGF-I in these cells. Morphological studies exhibited an increase in the lamellipodia formation upon IGF-I treatment, and confocal images of immunofluorescent staining revealed localization of phosphorylated FAK, paxillin, and vinculin at focal adhesions as well as redistribution of actin microfilaments and formation of actin stress fibers inside the cell. Western blotting, using antiphosphotyrosine demonstrated proteins with molecular masses of 125 kDa (FAK) and 68 kDa (paxillin) present in the IGF-I-treated cells, which were lacking in the control groups. In conclusion, these findings suggest that IGF-I can stimulate lamellipodia formation and promote adhesion of trophoblast cells to extracellular matrix by activating their adhesion molecules that must be activated within the implantation window.
Systemic Distribution of Steroid Sulfatase and Estrogen Sulfotransferase in Human Adult and Fetal Tissues
Authors: Miki Yasuhiro , Nakata Taisuke , Suzuki Takashi , Darnel Andrew David , Moriya Takuya , Kaneko Chika , Hidaka Kumiko , Shiotsu Yukimasa , Kusaka Hideaki , Sasano Hironobu
Published: December 2002

Estrogens play a key role in various target tissues. Enzymes involved in the biosynthesis and metabolism of these sex steroids also regulate estrogenic actions in these tissues. Estrone sulfate (E1S) is a major circulating plasma estrogen that is converted into the biologically active estrogen, estrone (E1), by steroid sulfatase (STS). E1 is also sulfated and reverted into E1S by estrogen sulfotransferase (EST). These two enzymes have recently been shown to play important roles in the in situ estrogen actions of various sex steroid-dependent human tumors. However, the distribution of STS and EST in normal adult and fetal human tissues remains largely unknown. Therefore, in this study, in addition to examining the tissue distribution of both STS and EST mRNA in human adult and fetal tissues using RT followed by quantitative PCR, we studied the activity of these enzymes using 3H-labeled E1/E1S as substrates in the homogenates of various human adult tissues. We also examined the localization of STS and EST protein in human adult and fetal tissues using immunohistochemistry, and that of EST mRNA in the adult kidney using laser dissection microscopy and PCR. STS mRNA, enzyme activity, and immunoreactivity were either absent or detected at very low levels in all adult and fetal tissues examined in this study. EST mRNA expression, however, was detected in all of the tissues examined, except for adult spleen and pancreas. EST enzyme activities were consistent with those of mRNA expression in the great majority of the tissues examined. Marked EST immunoreactivity was detected in hepatocytes, adrenal gland (adult, zona fasciculate to the reticularis; fetus, fetal zone), and epithelial cells of the gastrointestinal tract, smooth muscle cells of the tunica media in aorta, Leydig cells of the testis, and syncytiotrophoblast of the placenta. Patterns of EST immunolocalization were similar between adult and fetal human tissues, but EST immunoreactivity was detected in the urinary tubules of adult kidney, whereas in the fetal kidney, it was localized in the interstitial cells surrounding the urinary tubules. In the adult kidney, the presence of EST mRNA was also confirmed in the cells of urinary tubules using laser dissection microscopy and RT-PCR.

Although the number of human tissues available for examination in this study was limited, our results suggest that between the enzymes involved in estrogen activation or inactivation, EST and not STS is the more widely expressed enzyme in various peripheral tissues in humans. We speculate that EST may play an important role in protecting peripheral tissues from possible excessive estrogenic effects.
Thyroid Acropachy: Report of 40 Patients Treated at a Single Institution in a 26-Year Period
Authors: Fatourechi Vahab , Ahmed Debra D. F. , Schwartz Kara M.
Published: December 2002

Thyroid acropachy is an extreme manifestation of autoimmune thyroid disease. It presents with digital clubbing, swelling of digits and toes, and periosteal reaction of extremity bones. It is almost always associated with ophthalmopathy and thyroid dermopathy. During a 26-yr period at our institution, of 178 patients with thyroid dermopathy, 40 had acropachy. Clubbing associated with thyroid dermopathy (pretibial myxedema) was seen in 35 patients. Clubbing usually was not a patient complaint and was noted only by clinical observers. Four of eight patients with hand and extremity radiographs had periosteal reaction. Seven had associated extremity and joint pain; this pain was absent at long-term follow-up. Half of the patients required systemic corticosteroid therapy, 53% required transantral or transfrontal orbital decompression for severe ophthalmopathy, and 18% had the elephantiasic form of dermopathy. Cigarette-smoking rates were 81% for women and 75% for men (mean, 28 pack-years). All 13 patients who had thyroid-stimulating Ig measurement had high titers. Long-term follow-up (median, 12.5 yr) revealed that acropachy was not a complaint in follow-up visits or questionnaires. The data suggest that thyroid acropachy is an indicator of severity of ophthalmopathy and dermopathy. It is a source of clinical concern only if dermopathy is persistent and severe.
Growth Hormone Blunts Protein Oxidation and Promotes Protein Turnover to a Similar Extent in Abdominally Obese and Normal-Weight Women
Authors: Buijs Madelon M. , Romijn Johannes A. , Burggraaf Jacobus , De Kam Marieke L. , Cohen Adam F. , Frölich Marijke , Stellaard Frans , Meinders A. Edo , Pijl Hanno
Published: December 2002

Abdominally obese individuals have reduced 24-h plasma GH concentrations. Their normal plasma IGF-I levels may reflect GH hypersensitivity. Alternatively, obesity-associated hyposomatotropism may cause less biological effect in target tissues. We therefore determined whole-body responsiveness to the anabolic effects of GH in abdominally obese (OB) and normal weight (NW) premenopausal women. A 1-h iv infusion of GH or placebo was randomly administered to six NW (body mass index, 21.1 ± 1.9 kg/m2) and six OB (body mass index, 35.5 ± 1.5 kg/m2) women in a cross-over design. Endogenous insulin, glucagon and GH secretion was suppressed by infusion of somatostatin. Whole-body protein turnover was measured using a 10-h infusion of [13C]-leucine. GH administration induced a similar plasma GH peak in NW and OB women (49.8 ± 10.4 vs. 45.1 ± 5.6 mU/liter). GH, compared with placebo infusion, increased nonoxidative leucine disposal, P < 0.0001) and endogenous leucine appearance (Ra, P = 0.0004) but decreased leucine oxidation (P = 0.0051). All changes were similar in both groups. Accordingly, whole-body GH responsiveness, defined as the maximum response of nonoxidative leucine disposal, leucine Ra, and oxidation per unit of GH, was not different in OB and NW women (0.25 ± 0.18 vs. 0.19 ± 0.17 μmol/kg·h, 0.21 ± 0.23 vs. 0.13 ± 0.17 μmol/kg·h, and −0.10 ± 0.08 vs. −0.08 ± 0.05 μmol/kg·h, respectively). These results indicated that whole-body tissue responsiveness to the net anabolic effect of GH is similar in OB and NW women. Hence, we inferred that hyposomatotropism may promote amino acid oxidation and blunt protein turnover in abdominal obesity. However, hyposomatotropism cannot account for all anomalous features of protein metabolism in abdominally obese humans.
Demonstration of Enhanced Potency of a Chimeric Somatostatin-Dopamine Molecule, BIM-23A387, in Suppressing Growth Hormone and Prolactin Secretion from Human Pituitary Somatotroph Adenoma Cells
Authors: Saveanu A. , Lavaque E. , Gunz G. , Barlier A. , Kim S. , Taylor J. E. , Culler M. D. , Enjalbert A. , Jaquet P.
Published: December 2002

In acromegaly, the combination of somatostatin (SS) and dopamine (DA) agonists has been shown to enhance suppression of GH secretion. In the present study, a new chimeric molecule, BIM-23A387, which selectively binds to the SS subtype 2 receptor (sst2; Ki = 0.10 nm) and to the DA D2 receptor (D2DR; Ki = 22.1 nm) was tested in cultures prepared from 11 human GH-secreting tumors for its ability to suppress GH and prolactin (PRL) secretion. The chimeric compound was compared with individual sst2 and D2DR agonists of comparable activity at the individual receptors. All tumors expressed both sst2 and D2DR mRNAs (0.8 ± 0.2 and 4.7 ± 0.7 copy/copy β-glucuronidase mRNA, respectively). In cell cultures from seven octreotide-sensitive tumors, the maximal inhibition of GH release induced by the individual sst2 and D2DR analogs and by BIM-23A387 was similar. However, the mean EC50 for GH suppression by BIM-23A387 (0.2 pm) was 50 times lower than that of the individual sst2 and D2DR analogs, either used individually or combined. Similar data were obtained in four tumors that were only partially responsive to octreotide. The inhibition of GH release by BIM-23A387 was only partially reversed by the D2R2 antagonist, sulpiride, or by the sst2 antagonist, BIM-23454. Only when both antagonists were combined was the GH suppressive effect of BIM-23A387 totally reversed. Finally, BIM-23A387 produced a mean 73 ± 6% inhibition of PRL in six mixed GH plus PRL tumors. These data demonstrate an enhanced potency of the chimeric molecule, BIM-23A387, in suppressing GH and PRL secretion from acromegalic tumors, which cannot be explained merely on the basis of binding affinity for SS and/or DA receptors.
Effects of Pioglitazone in Nondiabetic Patients with Arterial Hypertension: A Double-Blind, Placebo-Controlled Study
Authors: Füllert S. , Schneider F. , Haak E. , Rau H. , Badenhoop K. , Lübben G. , Usadel K.-H. , Konrad T.
Published: December 2002

Hypertension is often associated with insulin resistance, dyslipidemia and obesity, which indicate a prediabetic state and increased risk of cardiovascular disease. Pioglitazone treatment of patients with type 2 diabetes reduces insulin resistance and improves lipid profiles. The present double-blind placebo-controlled study is the first study to report effects of pioglitazone in non-diabetic patients with arterial hypertension. Following a one week run-in, 60 patients were randomized to receive either pioglitazone (45 mg/day) or placebo for 16 weeks. Insulin sensitivity (M-value) increased by 1.2 ± 1.7 mg/min/kg with pioglitazone compared with 0.4 ± 1.4 mg/min/kg (P = 0.022) with placebo. HOMA index was decreased (−22.5 ± 45.8) by pioglitazone but not by placebo (+0.8 ± 26.5; P < 0.001). Decreases in fasting insulin and glucose were significantly (P = 0.002 and P = 0.004, respectively) greater with pioglitazone than placebo. Body weight did not change significantly with either treatment. HDL-cholesterol was increased and apolipoprotein B was decreased to a significantly greater extent with pioglitazone. There was a significantly (P = 0.016) greater decrease from baseline in diastolic blood pressure with pioglitazone. These changes would suggest improved glucose metabolism and a possible reduction in risk of cardiovascular disease with pioglitazone treatment of non-diabetic patients with arterial hypertension.
11β-Hydroxysteroid Dehydrogenase Types 1 and 2: An Important Pharmacokinetic Determinant for the Activity of Synthetic Mineralo- and Glucocorticoids
Authors: Diederich Sven , Eigendorff Ekkehard , Burkhardt Patrick , Quinkler Marcus , Bumke-Vogt Christiane , Rochel Marina , Seidelmann Dieter , Esperling Peter , Oelkers Wolfgang , Bähr Volker
Published: December 2002

The 11β-hydroxysteroid dehydrogenase (11β-HSD) system plays a pivotal role in glucocorticoid (GC) and mineralocorticoid (MC) action. Although 11β-HSD activities are important determinants for the efficacy of synthetic MCs and GCs, corresponding pharmacokinetic data are scanty. Therefore, we characterized 11β-HSD profiles for a wide range of steroids often used in clinical practice. 11β-HSD1 and 11β-HSD2 were selectively examined in 1) human liver and kidney cortex microsomes, and 2) Chinese hamster ovarian cells stably transfected with 11β-HSD1 or 11β-HSD2 expression vectors. Both systems produced concordant evidence for the following conclusions.

Oxidation of steroids by 11β-HSD2 is diminished if they are fluorinated in position 6α or 9α (e.g. in dexamethasone) or methylated at 2α or 6α (in methylprednisolone) or 16α or 16β, by a methylene group at 16 (in prednylidene), methyloxazoline at 16, 17 (in deflazacort), or a 2-chlor configuration. Whereas the methyl groups also decrease reductase activity (steric effects), fluorination increases reductase activity (negative inductive effect), leading to a shift to reductase activity. This may explain the strong MC activity of 9α-fluorocortisol and should be considered in GC therapy directed to 11β-HSD2-expressing tissues (kidney, colon, and placentofetal unit). 11β-HSD2 oxidation of prednisolone is more effective than that of cortisol, explaining the reduced MC activity of prednisolone compared with cortisol.

Reduction by 11β-HSD1 is diminished by 16α-methyl, 16β-methyl, 2α-methyl, and 2-chlor substitution, whereas it is increased by the Δ1-dehydro configuration in prednisone, resulting in higher hepatic first pass activation of prednisone compared with cortisone.

To characterize a GC or a MC as substrate for the different 11βHSDs may be essential for an optimized steroid therapy.
Phosphoinositide-3-kinase-independent contractile activities associated with Fcγ-receptor-mediated phagocytosis and macropinocytosis in macrophages
Authors: Araki Nobukazu , Hatae Tanenori , Furukawa Aizo , Swanson Joel A.
Published: December 2002

Previous studies have shown that Fcγ receptor (FcR)-mediated phagocytosis and macropinocytosis in macrophages consist of two dissociable activities: a phosphoinositide 3-kinase (PI3K)-independent extension of phagocytic cups and a PI3K-dependent contractile mechanism that closes phagosomes and ruffles into intracellular organelles. Here, we identify an additional contractile activity that persists in the presence of the PI3K inhibitor wortmannin. ML-7, an inhibitor of myosin-light-chain kinase (MLCK), inhibited FcR-mediated phagocytosis, macropinocytosis and cell movements associated with ruffling. Scanning electron microscopy demonstrated a striking difference in morphology between phagocytic cups in the different inhibitors: whereas phagocytic cups of control cells and wortmannin-treated cells conformed closely to particles and appeared to have constricted them, the phagocytic cups in cells treated with ML-7 were more open. Video microscopy of macrophages expressing green-fluorescent-protein (GFP)—actin fusions revealed that bound IgG-opsonized erythrocytes were squeezed during phagosome formation and closure. In ML-7, GFP—actin-rich protrusions extended outward but failed to squeeze particles. Moreover, in contrast to the effects of PI3K inhibitors, ML-7 markedly reduced ruffle movement, and perturbed circular ruffle formation. These PI3K-independent myosin-II-based contractile activities that squeeze phagocytic cups and curve ruffles therefore represent a third component activity of the actin cytoskeleton during phagocytosis and macropinocytosis.
Control of Texture in ZnO by Slip Casting in a Strong Magnetic Field and Heating
Authors: Suzuki Tohru S , Sakka Yoshio
Published: December 2002

Anisotropic susceptibilities of feeble magnetic ceramics such as zinc oxide are quite small; therefore, it had been difficult to control the textured microstructures of these ceramics using a magnetic field. However, fabrication of the textured ZnO is achieved by slip casting in a strong magnetic field followed by heating. After the stable suspension of ZnO fine particles was compacted by slip casting in a strong magnetic field, a green body with a slight degree of crystalline texture was obtained. Densification and the grain growth enhanced the degree of texture during the sintering.
Substrate and product inhibition of hydrogen production by the extreme thermophile, Caldicellulosiruptor saccharolyticus
Authors: van Niel Ed W. J. , Claassen Pieternel A. M. , Stams Alfons J. M.
Published: December 2002

Substrate and product inhibition of hydrogen production during sucrose fermentation by the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus was studied. The inhibition kinetics were analyzed with a noncompetitive, nonlinear inhibition model. Hydrogen was the most severe inhibitor when allowed to accumulate in the culture. Concentrations of 5–10 mM H2 in the gas phase (≡ partial hydrogen pressure (pH2) of (1–2) · 104 Pa) initiated a metabolic shift to lactate formation. The extent of inhibition by hydrogen was dependent on the density of the culture. The highest tolerance for hydrogen was found at low volumetric hydrogen production rates, as occurred in cultures with low cell densities. Under those conditions the critical hydrogen concentration in the gas phase was 27.7 mM H2 (≡ pH2 of 5.6 · 104 Pa); above this value hydrogen production ceased completely. With an efficient removal of hydrogen sucrose fermentation was mainly inhibited by sodium acetate. The critical concentrations of sucrose and acetate, at which growth and hydrogen production was completely inhibited (at neutral pH and 70°C), were 292 and 365 mM, respectively. Inorganic salts, such as sodium chloride, mimicked the effect of sodium acetate, implying that ionic strength was responsible for inhibition. Undissociated acetate did not contribute to inhibition of cultures at neutral or slightly acidic pH. Exposure of exponentially growing cultures to concentrations of sodium acetate or sodium chloride higher than ca. 175 mM caused cell lysis, probably due to activation of autolysins. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 255–262, 2003.
Effects of methanol concentration on expression levels of recombinant protein in fed‐batch cultures of Pichia methanolica
Authors: Mayson Brian E. , Kilburn Douglas G. , Zamost Bruce L. , Raymond Christopher K. , Lesnicki Gary J.
Published: December 2002

The methylotrophic yeast Pichia methanolica can be used to express recombinant genes at high levels under the control of the methanol‐inducible alcohol oxidase (AUG1) promoter. Methanol concentrations during the induction phase directly affect cellular growth and protein yield. Various methanol concentrations controlled by an on‐line monitoring and control system were investigated in mixed glucose/methanol fed‐batch cultures of P. methanolica expressing the human transferrin N‐lobe protein. The PMAD18 P. methanolica strain utilized is a knock‐out for the chromosomal AUG1 gene locus, resulting in a slow methanol utilization phenotype. Maximum growth of 100 g of dry cell weight per liter of culture was observed in cultures grown at 1.0% (v/v) methanol concentration. Maximum recombinant gene expression was observed for cultures controlled at 0.7% (v/v) methanol concentration, resulting in maximum volumetric production of 450 mg of transferrin per liter after 72 h of elapsed fermentation time. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 291–298, 2003.
Kinetic analysis of bacterial bioluminescence
Authors: Kelly Christine J. , Hsiung Chia‐Joung , Lajoie Curtis A.
Published: December 2002

Bioluminescence from the lux‐based bacterial reporter Pseudomonas fluorescens HK44 was experimentally investigated under growth substrate‐rich and limiting conditions in batch, continuous stirred tank (CSTR), and turbidostat reactors. A mechanistically based, mathematical model was developed to describe bioluminescence based on 1) production and decay of catalytic enzymes, and 2) reactant cofactor availability. In the model, bioluminescence was a function of inducer, growth substrate, and biomass concentration. A saturational dependence on growth substrate concentration accommodated dependence on cofactor availability and inducer concentration to accommodate enzyme production was incorporated in the model. Under growth substrate and inducer limiting conditions in the batch reactor and CSTR, bioluminescence was found to decrease in response to cellular energy limitations. The effective lux system enzyme decay rate was determined in independent measurements to be 0.35 hr−1 and the model captured most of the bioluminescent behavior, except at long growth times and high cell density. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 370–378, 2003.